N-hydroxy bicyclic hydantoin carbamates as tools for identification of serine hydrolase targets

ABSTRACT

Provided herein are N-hydroxy bicyclic hydantoin carbamates and pharmaceutical compositions comprising said compounds. The subject compounds and compositions are useful as modulators of serine hydrolases. Furthermore, the subject compounds and compositions are useful for the treatment of one or more of cancer, pain, diabetes, obesity/metabolic syndrome, epilepsy, traumatic brain injury, and inflammation.

CROSS-REFERENCE

This application is a U.S. National State Entry of PCT/US2015/031838,filed on May 20, 2015; which claims the benefit of priority from U.S.Provisional Application No. 62/001,869, filed May 22, 2014, all of whichare incorporated herein by reference in its entirety.

STATEMENT OF GOVERNMENT SUPPORT

This invention was made with government support under DA033760 andMH084512 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

BACKGROUND

Serine hydrolase inhibitors, which facilitate enzyme function assignmentand are used to treat a range of human disorders, often act by anirreversible mechanism that involves covalent modification of the serinehydrolase catalytic nucleophile.

BRIEF SUMMARY OF THE INVENTION

This disclosure is directed, in various embodiments, to a compoundhaving modulatory bioactivity with respect to a serine hydrolase.

One embodiment provides a compound of Formula (I):

wherein

-   -   W is

-   -   each m is independently 0, 1, or 2;    -   Y is N or CH;    -   R¹ is H, halo, —OH, cyano, amino, substituted or unsubstituted        (C₁-C₈)alkyl, substituted or unsubstituted (C₁-C₈)alkoxy,        substituted or unsubstituted (C₆-C₁₀)aryl, or substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl;    -   R² is H, —COOR⁵, or —CONR⁵R⁶;    -   R³ is H, substituted or unsubstituted (C₆-C₁₀)aryl, substituted        or unsubstituted 5-9 membered heteroaryl, substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, or substituted or        unsubstituted 5-9 membered heteroaryl(C₁-C₈)alkyl;    -   R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl, substituted or        unsubstituted 5-9 membered heteroaryl, substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or        unsubstituted 5-9 membered heteroaryl(C₁-C₈)alkyl, substituted        or unsubstituted di(C₆-C₁₀)aryl(C₁-C₈)alkyl;    -   R⁵ is H or substituted or unsubstituted (C₁-C₈)alkyl;    -   R⁶ is substituted or unsubstituted (C₆-C₁₀)aryl, substituted or        unsubstituted 5-9 membered heteroaryl, substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, or substituted or        unsubstituted 5-9 membered heteroaryl(C₁-C₈)alkyl;    -   Ring A is a 5-7 membered heterocyclyl containing 0-2 NR′,        wherein:    -   each R′ is independently H, substituted or unsubstituted        (C₁-C₈)alkyl, substituted or unsubstituted (C₁-C₈)alkoxy,        substituted or unsubstituted (C₁-C₈)alkylcarbonyl, substituted        or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted or        unsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted        (C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted        (C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted        (C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted        (C₆-C₁₀)aryl, substituted or unsubstituted (C₆-C₁₀)arylcarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryloxycarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl,        substituted or unsubstituted 5-9 membered heterocyclyl,        substituted or unsubstituted 5-9 membered        heterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted        5-9-membered heterocyclylcarbonyl, substituted or unsubstituted        5-9 membered heteroaryl, substituted or unsubstituted 5-9        membered heteroaryl(C₁-C₈)alkyl, or substituted or unsubstituted        5-9 membered heteroarylcarbonyl.

Another embodiment provides a pharmaceutical composition comprising anN-hydroxy bicyclic hydantoin carbamate described herein and at least onepharmaceutically acceptable excipient.

Another embodiment provides a method of modulation of a serinehydrolase, comprising contacting the serine hydrolase with an effectiveamount or concentration of an N-hydroxy bicyclic hydantoin carbamatedescribed herein.

Another embodiment provides a method of treatment of a medical conditionin a patient, wherein modulation of a serine hydrolase is medicallyindicated, comprising administering an effective dose of an N-hydroxybicyclic hydantoin carbamate described herein.

In some embodiments, the medical condition is selected from cancer,pain, diabetes, obesity/metabolic syndrome, epilepsy, traumatic braininjury, and inflammation.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows potency and selectivity of N-hydroxy bicyclic hydantoincarbamates (1-20) as assessed by gel-based competitive ABPP with theSH-specific ABPP probe FP-Rh in the mouse brain membrane proteome.

FIG. 2A shows in vitro IC₅₀ curves for LYPLA1/2 for Compound 1 inendogenous human LYPLA1/2 in a HeLa soluble proteome.

FIG. 2B shows in situ IC₅₀ curves for LYPLA1/2 for Compound 1 inendogenous human LYPLA1/2 in a in cultured HeLa cells.

FIG. 2C shows in vitro IC₅₀ curves for Compound 1 against endogenousmouse LYPLA1/2 and anti-targets ABHD6 and FAAH in a mouse brain membraneproteome.

FIG. 3A shows In situ inhibition of LYPLA1 and LYPLA2 by Compound 1.

FIG. 3B shows potent and selective in vivo activity for Compound 1.

FIG. 4A shows the inhibition profile of Compound 1 (1 μM, 2 hour in situtreatment, serum-supplemented medium)

FIG. 5 shows cytotoxicity analysis of Compound 1 and Compound 2 inserum-free and serum-supplemented medium.

FIG. 6A shows gel filtration studies revealing Compound 1 is anirreversible inhibitor.

FIG. 7A-1 shows Compounds 1, 2, 5, 7, 13, 21, 23, 31, and 38 in vitroinhibition profiles in a mouse brain membrane proteome.

FIG. 7A-2 shows ABPP-SILAC analysis to identify SH targets of Compound5.

FIG. 7B shows concentration-dependent inhibition of mouse brain membraneSHs by MJN193 (4-isopropyl-2,5-dioxoimidazolidin-1-yl4-(4-methoxyphenyl)piperazine-1-carboxylate), Compounds 1 and 38, asmeasured by gel-based competitive ABPP.

FIG. 8A shows recombinant expression of human ABHD3 and ABHD4 bytransient transfection in HEK293T cells and detection of both enzymes ina combined lysate of transfected cells by gel-based ABPP with the FP-Rhprobe.

FIG. 8B shows concentration-dependent inhibition of ABHD3 and ABHD4 byCompounds 13, 38, 35, 31, 36, and 37 using gel-based competitive ABPP.

FIG. 8C shows concentration-dependent inhibition of ABHD3 and ABHD4 byCompounds 13, 38, 35, 31, 36, and 37. Data represent competitivegel-based ABPP signals for recombinant ABHD3 and ABHD4 expressed bytransient transfection in HEK293T cells.

FIG. 9 shows ABPP-SILAC analysis showing overlaid in situ SH inhibitionprofiles for Compounds 38 (100 nM, 4 h) and 37 (1 μM, 4 h) in PC3 cells.Data represent average values±S.D. for two independent experiments.

FIG. 10 shows the reactivity with recombinant ABHD4 in transfected celllysates of Compound 39. Left side of gel shows a competitive ABPP gel ofABHD4-transfected cell lysates treated with the indicated concentrationsof Compound 39 (30 min 37° C.) followed by FP-Rh. Right side of gelshows direct labeling of ABHD4 and other SHs by Compound 39 as measuredby CuAAC to an Rh-N3 tag.

FIG. 11 shows Compound 39 detecting ABHD4 in mouse brain and testistissues. ABHD4+/+ and −/− tissues were treated with FP-Rh (1 μM, 30 min)or Compound 39 (1 μM, 60 min) and then analyzed by gel-based ABPP. A 37kDa band matching the predicted molecular mass of ABHD4 was detected byboth probes in ABHD4+/+, but not ABHD4−/− tissues.

FIG. 12 shows ABPP-SILAC analysis showing the in situ SH inhibitionprofile for Compound 37 (1 μM, 4 h) in PC3 cells, where SH enrichmentand inhibition were measured with the Compound 39 probe. Data representaverage values±S.D. for two independent experiments.

FIG. 13 shows ABPP-SILAC analysis comparing the reactivity andenrichment of SHs with FP-biotin (2.5 μM, 1 h) versus Compound 39 (5 μM,1 h) in PC3 cell proteomes. Data represent average values±S.D. for twoindependent experiments.

FIG. 14 shows the verification that recombinant human PPT1 expressed bytransient transfection in HEK293T cells reacts with Compound 39 (1 μM),but not FP-Rh (1 μM), and the Compound 39 reactivity is blocked byCompound 37 (5 μM, 4 h).

FIG. 15 shows that Compound 39 labels recombinant wild type PPT1, butnot the catalytic serine mutant (S115A) of this enzyme.

FIG. 16 shows an isoTOP-ABPP experiment identifying the catalytic serineS115 as the site of Compound 39 labeling in PPT1. Shown is the structureof the Compound 39-PPT1 adduct (connected by CuAAC to the azide-tag usedto enrich and release Compound 39-reactive peptides in isoTOP-ABPPexperiments).

FIG. 17A shows that hPPT1-transfected cell lysates show much greaterhydrolytic activity with the 4-methylumbelliferyl oleate (4-MUBO)substrate compared to mock-transfected cell lysates or hPPT1-transfectedcell lysates pre-treated with Compound 26 (50 μM, 30 min). Datarepresent average values±S.E. for three independent experiments.

FIG. 17B shows concentration-dependent inhibition of 4-MUBO hydrolyticactivity of hPPT1-transfected cell lysates by Compounds 26, 34, and 1.Data represent average values±S.E. for three independent experiments.

FIG. 17C shows that PC3 cells treated in situ with Compound 26, but notCompound 34 (500 nM inhibitor; 4 h) showed substantial reductions in4-MUBO hydrolysis activity compared to DMSO-treated control cells. Datarepresent average values±S.E. for three independent experiments.

FIG. 17D shows the screening of MJN193(4-isopropyl-2,5-dioxoimidazolidin-1-yl4-(4-methoxyphenyl)piperazine-1-carboxylate), Compounds 1, 5, 13, 34,37, 24, 25, and 26 in hPPT1-transfected HEK293T cells in situ, wherePPT1 inhibition was measured by competitive ABPP using the Compound 39probe.

FIG. 18 shows that ABHD4-transfected HEK293T cell lysates show muchgreater NAPE lipase activity compared to mock-transfected HEK239T celllysates (top) and this activity is inhibited by N-hydroxy bicyclichydantoin carbamates (bottom). NAPE hydrolysis was measured by followingthe release of oleic acid from the NAPE substrate(1,2-dioleoyl-sn-glycero-3-phospho (N-arachidonoyl) ethanolamine).

FIG. 19 shows ABPP-SILAC analysis showing overlaid in situ SH inhibitionprofiles for Compound 26 (0.1 μM, 4 h) and Compound 34 (1 μM, 4 h) inPC3 cells where SH enrichment and inhibition were measured with theFP-biotin (top) or Compound 39 (bottom) probes. Data represent averagevalues±S.D. for two independent experiments.

FIG. 20 shows ABPP-SILAC analysis showing in situ SH inhibition profilesfor Compound 26 (1 μM, 4 h) in PC3 cells where SH enrichment andinhibition were measured with the Compound 39 probe in Compound 26-versus DMSO-treated cells. Data represent average values±S.D. for twoindependent experiments.

FIG. 21 shows gel-based competitive ABPP analysis of SH activities intissues from Compound 26 treated mice. The activities of ABHD4 and PPT1were visualized using the Compound 39 probe in tissue proteomes treatedwith PNGaseF (post-Compound 39 labeling), while the activities of LYPLA1and LYPLA2 were visualized using the FP-Rh probe.

FIG. 22 shows full gels showing competitive ABPP results for tissuesfrom mice treated with Compound 26 or Compound 1. The left and rightgels in each pair represent tissues treated with the FP-Rh (1 μM, 30min) and Compound 39 (2 μM, 60 min), respectively.

FIG. 23 shows gel-based competitive ABPP analysis of SH activities intissues from Compound 1 treated mice. The activities of ABHD4 and PPT1were visualized using the Compound 39 probe in tissue proteomes treatedwith PNGaseF (post-Compound 39 labeling), while the activities of LYPLA1and LYPLA2 were visualized using the FP-Rh probe.

FIG. 24 shows ABPP-SILAC analysis showing in situ SH inhibition profilesfor Compound 1 (1 μM, 4 h) in PC3 cells where SH enrichment andinhibition were measured with the FP-biotin probe in Compound 1 versusDMSO-treated cells. Data represent average values±S.D. for twoindependent experiments.

DETAILED DESCRIPTION OF THE INVENTION

This disclosure is directed, in various embodiments, to a compoundhaving modulatory bioactivity with respect to a serine hydrolase.

Serine hydrolases (SHs) represent one of the largest and most diverseenzyme classes in Nature and perform myriad biochemical functions inphysiology and disease. SHs use a conserved mechanism involving abase-activated serine nucleophile to hydrolyze amide, ester, andthioester bonds in biomolecules; however, these enzymes also displaymarkedly different structures and folds, distribute across virtually allsubcellular compartments in the cell, and accept an expansive array ofsmall- and macro-molecule substrates. In accordance with their diversebiological activities, SHs are targeted by drugs that are used to treata wide range of diseases, including cognitive dementia, obesity,diabetes, and bacterial and viral infections.

As used herein and in the appended claims, the singular forms “a,”“and,” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “an agent” includesa plurality of such agents, and reference to “the cell” includesreference to one or more cells (or to a plurality of cells) andequivalents thereof. When ranges are used herein for physicalproperties, such as molecular weight, or chemical properties, such aschemical formulae, all combinations and subcombinations of ranges andspecific embodiments therein are intended to be included. The term“about” when referring to a number or a numerical range means that thenumber or numerical range referred to is an approximation withinexperimental variability (or within statistical experimental error), andthus the number or numerical range varies between 1% and 15% of thestated number or numerical range. The term “comprising” (and relatedterms such as “comprise” or “comprises” or “having” or “including”) isnot intended to exclude that which in other certain embodiments, forexample, an embodiment of any composition of matter, composition,method, or process, or the like, described herein, may “consist of” or“consist essentially of” the described features.

Definitions

As used in the specification and appended claims, unless specified tothe contrary, the following terms have the meaning indicated below.

“Amino” refers to the —NH₂ radical.

“Carbonyl” refers to the >C═O radical.

“Cyano” refers to the —CN radical.

“Nitro” refers to the —NO₂ radical.

“Oxa” refers to the —O— radical.

“Oxo” refers to the ═O radical.

“Thioxo” refers to the ═S radical.

“Imino” refers to the ═N−H radical.

“Oximo” refers to the ═N—OH radical.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to fifteen carbon atoms (e.g.,(C₁-C₁₅)alkyl). In certain embodiments, an alkyl comprises one tothirteen carbon atoms (e.g., (C₁-C₁₃)alkyl). In certain embodiments, analkyl comprises one to eight carbon atoms (e.g., (C₁-C₈)alkyl). In otherembodiments, an alkyl comprises one to five carbon atoms (e.g.,(C₁-C₅)alkyl). In other embodiments, an alkyl comprises one to fourcarbon atoms (e.g., (C₁-C₄)alkyl). In other embodiments, an alkylcomprises one to three carbon atoms (e.g., (C₁-C₃)alkyl). In otherembodiments, an alkyl comprises one to two carbon atoms (e.g.,(C₁-C₂)alkyl). In other embodiments, an alkyl comprises one carbon atom(e.g., (C₁)alkyl). In other embodiments, an alkyl comprises five tofifteen carbon atoms (e.g., (C₅-C₁₅)alkyl). In other embodiments, analkyl comprises five to eight carbon atoms (e.g., (C₅-C₈)alkyl). Inother embodiments, an alkyl comprises two to five carbon atoms (e.g.,(C₂-C₅)alkyl). In other embodiments, an alkyl comprises three to fivecarbon atoms (e.g., (C₃-C₅)alkyl). In other embodiments, the alkyl groupis selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl(iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl),2-methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl(n-pentyl). The alkyl is attached to the rest of the molecule by asingle bond. Unless stated otherwise specifically in the specification,an alkyl group is optionally substituted by one or more of the followingsubstituents: halo, cyano, nitro, oxo, thioxo, imino, oximo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(f),—OC(O)—NR^(a)R^(f), —N(R^(a))C(O)R^(f), —N(R^(a))S(O)_(t)R^(f) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(f) (where tis 1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a)is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl or heteroarylalkyl, and each R^(f) is independently alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Alkoxy” refers to a radical bonded through an oxygen atom of theformula —O-alkyl, where alkyl is an alkyl chain as defined above.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon double bond, and having from two to twelvecarbon atoms. In certain embodiments, an alkenyl comprises two to eightcarbon atoms. In certain embodiments, an alkenyl comprises two to sixcarbon atoms. In other embodiments, an alkenyl comprises two to fourcarbon atoms. The alkenyl is attached to the rest of the molecule by asingle bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e.,allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. Unlessstated otherwise specifically in the specification, an alkenyl group isoptionally substituted by one or more of the following substituents:halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(f), —OC(O)—NR^(a)R^(f),—N(R^(a))C(O)R^(f), —N(R^(a))S(O)_(t)R^(f) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(f) (where t is 1 or 2)and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl or heteroarylalkyl, and each R^(f) is independently alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon triple bond, having from two to twelve carbonatoms. In certain embodiments, an alkynyl comprises two to eight carbonatoms. In certain embodiments, an alkynyl comprises two to six carbonatoms. In other embodiments, an alkynyl has two to four carbon atoms.The alkynyl is attached to the rest of the molecule by a single bond,for example, ethynyl, propynyl, butyryl, pentynyl, hexynyl, and thelike. Unless stated otherwise specifically in the specification, analkynyl group is optionally substituted by one or more of the followingsubstituents: halo, cyano, nitro, oxo, thioxo, imino, oximo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(f),—OC(O)—NR^(a)R^(f), —N(R^(a))C(O)R^(f), —N(R^(a))S(O)_(t)R^(f) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(f) (where tis 1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a)is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl or heteroarylalkyl, and each R^(f) is independently alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon chain linking the rest of the molecule to a radical group,consisting solely of carbon and hydrogen, containing no unsaturation andhaving from one to twelve carbon atoms, for example, methylene,ethylene, propylene, n-butylene, and the like. The alkylene chain isattached to the rest of the molecule through a single bond and to theradical group through a single bond. The points of attachment of thealkylene chain to the rest of the molecule and to the radical group arethrough one carbon in the alkylene chain or through any two carbonswithin the chain. In certain embodiments, an alkylene comprises one toeight carbon atoms (e.g., (C₁-C₈)alkylene). In other embodiments, analkylene comprises one to five carbon atoms (e.g., (C₁-C₅)alkylene). Inother embodiments, an alkylene comprises one to four carbon atoms (e.g.,(C₁-C₄)alkylene). In other embodiments, an alkylene comprises one tothree carbon atoms (e.g., (C₁-C₃)alkylene). In other embodiments, analkylene comprises one to two carbon atoms (e.g., (C₁-C₂)alkylene). Inother embodiments, an alkylene comprises one carbon atom (e.g.,(C₁)alkylene). In other embodiments, an alkylene comprises five to eightcarbon atoms (e.g., (C₅-C₈)alkylene). In other embodiments, an alkylenecomprises two to five carbon atoms (e.g., (C₂-C₅)alkylene). In otherembodiments, an alkylene comprises three to five carbon atoms (e.g.,(C₃-C₅)alkylene). Unless stated otherwise specifically in thespecification, an alkylene chain is optionally substituted by one ormore of the following substituents: halo, cyano, nitro, oxo, thioxo,imino, oximo, trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(f), —OC(O)—NR^(a)R^(f), —N(R^(a))C(O)R^(f),—N(R^(a))S(O)_(t)R^(f) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)R^(f) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl, and each R^(f) isindependently alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl,aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Aryl” refers to a radical derived from an aromatic monocyclic ormulticyclic hydrocarbon ring system by removing a hydrogen atom from aring carbon atom. The aromatic monocyclic or multicyclic hydrocarbonring system contains only hydrogen and carbon from five to eighteencarbon atoms (e.g., (C₅-C₁₈)aryl), where at least one of the rings inthe ring system is fully unsaturated, i.e., it contains a cyclic,delocalized (4n+2) π-electron system in accordance with the Hückeltheory. In certain embodiments, an aryl comprises six to ten carbonatoms (e.g., (C₆-C₁₀)aryl). In certain embodiments, an aryl comprisessix carbon atoms (e.g., (C₆)aryl). The ring system from which arylgroups are derived include, but are not limited to, groups such asbenzene, fluorene, indane, indene, tetralin and naphthalene. Unlessstated otherwise specifically in the specification, the term “aryl” orthe prefix “ar-” (such as in “aralkyl”) is meant to include arylradicals optionally substituted by one or more substituentsindependently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)R^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl (optionally substituted with one or more halo groups), aralkyl,heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, eachR^(b) is independently a direct bond or a straight or branched alkyleneor alkenylene chain, and R^(c) is a straight or branched alkylene oralkenylene chain, and where each of the above substituents isunsubstituted unless otherwise indicated.

“Aryloxy” refers to a radical bonded through an oxygen atom of theformula —O-aryl, where aryl is as defined above.

“Aralkyl” or “arylalkyl” refers to a radical of the formula —R^(c)-arylwhere R^(c) is an alkylene chain as defined above, for example,methylene, ethylene, and the like. The alkylene chain part of thearalkyl radical is optionally substituted as described above for analkylene chain. The aryl part of the aralkyl radical is optionallysubstituted as described above for an aryl group. In some embodiments,the aralkyl is described as (C₆-C₁₀)aryl(C₁-C₈)alkyl where the(C₆-C₁₀)aryl and (C₁-C₈)alkyl are as defined above.

“Aralkenyl” refers to a radical of the formula —R^(d)-aryl where R^(d)is an alkenylene chain as defined above. The aryl part of the aralkenylradical is optionally substituted as described above for an aryl group.The alkenylene chain part of the aralkenyl radical is optionallysubstituted as defined above for an alkenylene group.

“Aralkynyl” refers to a radical of the formula —R^(e)-aryl, where R^(e)is an alkynylene chain as defined above. The aryl part of the aralkynylradical is optionally substituted as described above for an aryl group.The alkynylene chain part of the aralkynyl radical is optionallysubstituted as defined above for an alkynylene chain.

“Carbocyclyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,which includes fused or bridged ring systems, having from three tofifteen carbon atoms. In certain embodiments, a carbocyclyl comprisesthree to ten carbon atoms. In other embodiments, a carbocyclyl comprisesfive to seven carbon atoms. The carbocyclyl is attached to the rest ofthe molecule by a single bond. Carbocyclyl is saturated, (i.e.,containing single C—C bonds only) or unsaturated (i.e., containing oneor more double bonds or triple bonds). A fully saturated carbocyclylradical is also referred to as “cycloalkyl.” Examples of monocycliccycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl. In certain embodiments, acycloalkyl comprises three to eight carbon atoms (e.g.,(C₃-C₈)cycloalkyl). In other embodiments, a cycloalkyl comprises threeto seven carbon atoms (e.g., (C₃-C₇)cycloalkyl). In other embodiments, acycloalkyl comprises three to six carbon atoms (e.g.,(C₃-C₆)cycloalkyl). In other embodiments, a cycloalkyl comprises threeto five carbon atoms (e.g., (C₃-C₅)cycloalkyl). In other embodiments, acycloalkyl comprises three to four carbon atoms (e.g.,(C₃-C₄)cycloalkyl). An unsaturated carbocyclyl is also referred to as“cycloalkenyl.” Examples of monocyclic cycloalkenyls include, e.g.,cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycycliccarbocyclyl radicals include, for example, adamantyl, norbornyl (i.e.,bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl,7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwisestated specifically in the specification, the term “carbocyclyl” ismeant to include carbocyclyl radicals that are optionally substituted byone or more substituents independently selected from alkyl, alkenyl,alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionallysubstituted aryl, optionally substituted aralkyl, optionally substitutedaralkenyl, optionally substituted aralkynyl, optionally substitutedcarbocyclyl, optionally substituted carbocyclylalkyl, optionallysubstituted heterocyclyl, optionally substituted heterocyclylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, —R^(b)—OR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a),—R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)R^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl orheteroarylalkyl, each R^(b) is independently a direct bond or a straightor branched alkylene or alkenylene chain, and R^(c) is a straight orbranched alkylene or alkenylene chain, and where each of the abovesubstituents is unsubstituted unless otherwise indicated.

“Carbocyclylalkyl” refers to a radical of the formula —R^(c)-carbocyclylwhere R^(c) is an alkylene chain as defined above. The alkylene chainand the carbocyclyl radical is optionally substituted as defined above.In some embodiments, the carbocyclylalkyl is described as(C₃-C₈)carbocyclyl(C₁-C₈)alkyl where the (C₃-C₈)carbocyclyl and(C₁-C₈)alkyl are as defined above.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodosubstituents.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, fluoromethyl,2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. Insome embodiments, the alkyl part of the fluoroalkyl radical isoptionally substituted as defined above for an alkyl group.

“Heterocyclyl” refers to a stable 3- to 18-membered non-aromatic ringradical that comprises two to twelve carbon atoms and from one to sixheteroatoms selected from nitrogen, oxygen and sulfur. Unless statedotherwise specifically in the specification, the heterocyclyl radical isa monocyclic, bicyclic, tricyclic or tetracyclic ring system, whichincludes fused or bridged ring systems. The heteroatoms in theheterocyclyl radical are optionally oxidized. One or more nitrogenatoms, if present, are optionally quaternized. The heterocyclyl radicalis partially or fully saturated. In some embodiments, the heterocyclylis attached to the rest of the molecule through any atom of the ring(s).Examples of such heterocyclyl radicals include, but are not limited to,dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in thespecification, the term “heterocyclyl” is meant to include heterocyclylradicals as defined above that are optionally substituted by one or moresubstituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,oxo, thioxo, cyano, nitro, optionally substituted aryl, optionallysubstituted aralkyl, optionally substituted aralkenyl, optionallysubstituted aralkynyl, optionally substituted carbocyclyl, optionallysubstituted carbocyclylalkyl, optionally substituted heterocyclyl,optionally substituted heterocyclylalkyl, optionally substitutedheteroaryl, optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)R^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl orheteroarylalkyl, each R^(b) is independently a direct bond or a straightor branched alkylene or alkenylene chain, and R^(c) is a straight orbranched alkylene or alkenylene chain, and where each of the abovesubstituents is unsubstituted unless otherwise indicated.

“Heterocyclylalkyl” refers to a radical of the formula—R^(c)-heterocyclyl where R^(c) is an alkylene chain as defined above.If the heterocyclyl is a nitrogen-containing heterocyclyl, theheterocyclyl is optionally attached to the alkyl radical at the nitrogenatom. The alkylene chain of the heterocyclylalkyl radical is optionallysubstituted as defined above for an alkylene chain. The heterocyclylpart of the heterocyclylalkyl radical is optionally substituted asdefined above for a heterocyclyl group.

“Heterocyclylalkyl” refers to a radical of the formula—R^(c)-heterocyclyl where R^(c) is an alkylene chain as defined above.The alkylene chain and the carbocyclyl radical is optionally substitutedas defined above. In some embodiments, the heterocyclylalkyl isdescribed as (C₂-C₈)heterocyclyl(C₁-C₈)alkyl where the(C₂-C₈)heterocyclyl and (C₁-C₈)alkyl are as defined above.

“Heterocyclylalkoxy” refers to a radical bonded through an oxygen atomof the formula —O—R^(c)-heterocyclyl where R^(c) is an alkylene chain asdefined above. If the heterocyclyl is a nitrogen-containingheterocyclyl, the heterocyclyl is optionally attached to the alkylradical at the nitrogen atom. The alkylene chain of theheterocyclylalkoxy radical is optionally substituted as defined abovefor an alkylene chain. The heterocyclyl part of the heterocyclylalkoxyradical is optionally substituted as defined above for a heterocyclylgroup.

“Heteroaryl” refers to a radical derived from a 3- to 18-memberedaromatic ring radical that comprises two to seventeen carbon atoms(e.g., (C₂-C₁₈)heteroaryl) and from one to six heteroatoms selected fromnitrogen, oxygen and sulfur. As used herein, the heteroaryl radical is amonocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein atleast one of the rings in the ring system is fully unsaturated, i.e., itcontains a cyclic, delocalized (4n+2) π-electron system in accordancewith the Hückel theory. Heteroaryl includes fused or bridged ringsystems. The heteroatom(s) in the heteroaryl radical is optionallyoxidized. One or more nitrogen atoms, if present, are optionallyquaternized. The heteroaryl is attached to the rest of the moleculethrough any atom of the ring(s). Examples of heteroaryls include, butare not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl,1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl,benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl,1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl,benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl,benzofuranonyl, benzothienyl (benzothiophenyl),benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl,pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl,pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl,quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, theterm “heteroaryl” is meant to include heteroaryl radicals as definedabove which are optionally substituted by one or more substituentsselected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl,haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl,optionally substituted aralkyl, optionally substituted aralkenyl,optionally substituted aralkynyl, optionally substituted carbocyclyl,optionally substituted carbocyclylalkyl, optionally substitutedheterocyclyl, optionally substituted heterocyclylalkyl, optionallysubstituted heteroaryl, optionally substituted heteroarylalkyl,—R^(b)—OR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a),—R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)R^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl orheteroarylalkyl, each R^(b) is independently a direct bond or a straightor branched alkylene or alkenylene chain, and R^(c) is a straight orbranched alkylene or alkenylene chain, and where each of the abovesubstituents is unsubstituted unless otherwise indicated.

“N-heteroaryl” refers to a heteroaryl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heteroaryl radical to the rest of the molecule is through a nitrogenatom in the heteroaryl radical. An N-heteroaryl radical is optionallysubstituted as described above for heteroaryl radicals.

“C-heteroaryl” refers to a heteroaryl radical as defined above and wherethe point of attachment of the heteroaryl radical to the rest of themolecule is through a carbon atom in the heteroaryl radical. AC-heteroaryl radical is optionally substituted as described above forheteroaryl radicals.

“Heteroaryloxy” refers to radical bonded through an oxygen atom of theformula —O-heteroaryl, where heteroaryl is as defined above.

“Heteroarylalkyl” refers to a radical of the formula —R^(c)-heteroaryl,where R^(c) is an alkylene chain as defined above. If the heteroaryl isa nitrogen-containing heteroaryl, the heteroaryl is optionally attachedto the alkyl radical at the nitrogen atom. The alkylene chain of theheteroarylalkyl radical is optionally substituted as defined above foran alkylene chain. The heteroaryl part of the heteroarylalkyl radical isoptionally substituted as defined above for a heteroaryl group. In someembodiments, the heteroarylalkyl is described as(C₂-C₁₀)heteroaryl(C₁-C₈)alkyl where the (C₂-C₁₀)heteroaryl and(C₁-C₈)alkyl are as defined above.

“Heteroarylalkoxy” refers to a radical bonded through an oxygen atom ofthe formula —O—R^(c)-heteroaryl, where R^(c) is an alkylene chain asdefined above. If the heteroaryl is a nitrogen-containing heteroaryl,the heteroaryl is optionally attached to the alkyl radical at thenitrogen atom. The alkylene chain of the heteroarylalkoxy radical isoptionally substituted as defined above for an alkylene chain. Theheteroaryl part of the heteroarylalkoxy radical is optionallysubstituted as defined above for a heteroaryl group.

In some embodiments, the compounds disclosed herein contain one or moreasymmetric centers and thus give rise to enantiomers, diastereomers, andother stereoisomeric forms that are defined, in terms of absolutestereochemistry, as (R)— or (S)—. Unless stated otherwise, it isintended that all stereoisomeric forms of the compounds disclosed hereinare contemplated by this disclosure. When the compounds described hereincontain alkene double bonds, and unless specified otherwise, it isintended that this disclosure includes both E and Z geometric isomers(e.g., cis or trans.) Likewise, all possible isomers, as well as theirracemic and optically pure forms, and all tautomeric forms are alsointended to be included. The term “geometric isomer” refers to E or Zgeometric isomers (e.g., cis or trans) of an alkene double bond. Theterm “positional isomer” refers to structural isomers around a centralring, such as ortho-, meta-, and para-isomers around a benzene ring.

A “tautomer” refers to a molecule wherein a proton shift from one atomof a molecule to another atom of the same molecule is possible. Incertain embodiments, the compounds presented herein exist as tautomers.In circumstances where tautomerization is possible, a chemicalequilibrium of the tautomers will exist. The exact ratio of thetautomers depends on several factors, including physical state,temperature, solvent, and pH. Some examples of tautomeric equilibriuminclude:

“Optional” or “optionally” means that a subsequently described event orcircumstance may or may not occur and that the description includesinstances when the event or circumstance occurs and instances in whichit does not. For example, “optionally substituted aryl” means that thearyl radical is or is not substituted and that the description includesboth substituted aryl radicals and aryl radicals having no substitution.“Optionally substituted” and “substituted or unsubstituted” areinterchangeable.

“Pharmaceutically acceptable salt” includes both acid and base additionsalts. A pharmaceutically acceptable salt of any one of the N-hydroxybicyclic hydantoin carbamates described herein is intended to encompassany and all pharmaceutically suitable salt forms. Preferredpharmaceutically acceptable salts of the compounds described herein arepharmaceutically acceptable acid addition salts and pharmaceuticallyacceptable base addition salts.

“Pharmaceutically acceptable acid addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid,hydrofluoric acid, phosphorous acid, and the like. Also included aresalts that are formed with organic acids such as aliphatic mono- anddicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoicacids, alkanedioic acids, aromatic acids, aliphatic and. aromaticsulfonic acids, etc. and include, for example, acetic acid,trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like. Exemplary salts thus include sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates,trifluoroacetates, propionates, caprylates, isobutyrates, oxalates,malonates, succinate suberates, sebacates, fumarates, maleates,mandelates, benzoates, chlorobenzoates, methylbenzoates,dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates,phenylacetates, citrates, lactates, malates, tartrates,methanesulfonates, and the like. Also contemplated are salts of aminoacids, such as arginates, gluconates, and galacturonates (see, forexample, Berge S. M. et al., “Pharmaceutical Salts,” Journal ofPharmaceutical Science, 66:1-19 (1997)). Acid addition salts of basiccompounds are prepared by contacting the free base forms with asufficient amount of the desired acid to produce the salt.

“Pharmaceutically acceptable base addition salt” refers to those saltsthat retain the biological effectiveness and properties of the freeacids, which are not biologically or otherwise undesirable. These saltsare prepared from addition of an inorganic base or an organic base tothe free acid. In some embodiments, pharmaceutically acceptable baseaddition salts are formed with metals or amines, such as alkali andalkaline earth metals or organic amines. Salts derived from inorganicbases include, but are not limited to, sodium, potassium, lithium,ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminumsalts and the like. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary, and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, for example,isopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol,2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine,caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine,hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline,N-methylglucamine, glucosamine, methylglucamine, theobromine, purines,piperazine, piperidine, N-ethylpiperidine, polyamine resins and thelike. See Berge et al., supra.

As used herein, “treatment” or “treating” or “palliating” or“ameliorating” are used interchangeably herein. These terms refers to anapproach for obtaining beneficial or desired results including but notlimited to therapeutic benefit and/or a prophylactic benefit. By“therapeutic benefit” is meant eradication or amelioration of theunderlying disorder being treated. Also, a therapeutic benefit isachieved with the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient, notwithstanding that thepatient is still afflicted with the underlying disorder. Forprophylactic benefit, the compositions are administered to a patient atrisk of developing a particular disease, or to a patient reporting oneor more of the physiological symptoms of a disease, even though adiagnosis of this disease has not been made.

Compounds

N-hydroxy bicyclic hydantoin carbamates are described herein which areinhibitors of serine hydrolase. These compounds, and compositionscomprising these compounds, are useful for the treatment of cancer,pain, diabetes, obesity/metabolic syndrome, epilepsy, traumatic braininjury, and/or inflammation.

One embodiment provides a compound of Formula (I):

wherein

-   -   W is

-   -   each m is independently 0, 1, or 2;    -   Y is N or CH;    -   R¹ is H, halo, —OH, cyano, amino, substituted or unsubstituted        (C₁-C₈)alkyl, substituted or unsubstituted (C₁-C₈)alkoxy,        substituted or unsubstituted (C₆-C₁₀)aryl, or substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl;    -   R² is H, —COOR⁵, or —CONR⁵R⁶;    -   R³ is H, substituted or unsubstituted (C₆-C₁₀)aryl, substituted        or unsubstituted 5-9-membered heteroaryl, substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, or substituted or        unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl;    -   R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl, substituted or        unsubstituted 5-9-membered heteroaryl, substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or        unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl, or        substituted or unsubstituted di(C₆-C₁₀)aryl(C₁-C₈)alkyl;    -   R⁵ is H or substituted or unsubstituted (C₁-C₈)alkyl;    -   R⁶ is substituted or unsubstituted (C₆-C₁₀)aryl, substituted or        unsubstituted 5-9-membered heteroaryl, substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, or substituted or        unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl;    -   Ring A is a 5-7-membered heterocyclyl containing 0-2 NR′,        wherein:    -   each R′ is independently H, substituted or unsubstituted        (C₁-C₈)alkyl, substituted or unsubstituted (C₁-C₈)alkoxy,        substituted or unsubstituted (C₁-C₈)alkylcarbonyl, substituted        or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted or        unsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted        (C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted        (C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted        (C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted        (C₆-C₁₀)aryl, substituted or unsubstituted (C₆-C₁₀)arylcarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryloxycarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl,        substituted or unsubstituted 5-9-membered heterocyclyl,        substituted or unsubstituted 5-9-membered        heterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted        5-9-membered heterocyclylcarbonyl, substituted or unsubstituted        5-9-membered heteroaryl, substituted or unsubstituted        5-9-membered heteroaryl(C₁-C₈)alkyl, or substituted or        unsubstituted 5-9-membered heteroarylcarbonyl.

In some embodiments of a compound of Formula (I), each m is 0. In someembodiments of a compound of Formula (I), one m is 0 and the other mis 1. In some embodiments of a compound of Formula (I), each m is 1. Insome embodiments of a compound of Formula (I), R¹ is H, halo, —OH,cyano, amino, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₆-C₁₀)aryl, or(C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), R¹ is H. In some embodiments of a compound of Formula (I), R¹ is For Cl. In some embodiments of a compound of Formula (I), R¹ is methyl ormethoxy. In some embodiments of a compound of Formula (I), R¹ is phenyl.In some embodiments of a compound of Formula (I), R¹ is benzyl.

In some embodiments of a compound of Formula (I), W is

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted (C₆-C₁₀)aryl, orsubstituted or unsubstituted 5-9-membered heteroaryl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted (C₆-C₁₀)aryl, orsubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted (C₆-C₁₀)aryl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted phenyl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is unsubstituted phenyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is phenyl substituted with one or more substituentsselected from halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl,(C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is phenyl substituted with one or more substituentsselected from fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy.In some embodiments of Formula (I), W is

R² is H; and R³ is mono-substituted phenyl. In some embodiments ofFormula (I), W is

R² is H; and R³ is di-substituted phenyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is methoxy substituted phenyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is propargyloxy substituted phenyl. In some embodimentsof a compound of Formula (I), W is

R² is H; and R³ is fluoro substituted phenyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is chloro substituted phenyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is ethynyl substituted phenyl.

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted 5-9-membered heteroaryl.In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted 5-9-membered heteroaryl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is unsubstituted 5-9-membered heteroaryl.

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), W is

R² is H; and R³ is substituted or unsubstituted benzyl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is unsubstituted benzyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is benzyl substituted with one or more substituentsselected from halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl,(C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is benzyl substituted with one or more substituentsselected from fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy.In some embodiments of Formula (I), W is

R² is H; and R³ is mono-substituted benzyl. In some embodiments ofFormula (I), W is

R² is H; and R³ is di-substituted benzyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is methoxy substituted benzyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is propargyloxy substituted benzyl. In some embodimentsof a compound of Formula (I), W is

R² is H; and R³ is fluoro substituted benzyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is chloro substituted benzyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is ethynyl substituted benzyl.

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), W is

R² is H; and R³ is substituted or unsubstituted phenethyl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is unsubstituted phenethyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is phenethyl substituted with one or more substituentsselected from halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl,(C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is phenethyl substituted with one or more substituentsselected from fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy.In some embodiments of Formula (I), W is

R² is H; and R³ is mono-substituted phenethyl. In some embodiments ofFormula (I), W is

R² is H; and R³ is di-substituted phenethyl. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is methoxy substituted phenethyl. In some embodiments ofa compound of Formula (I), W is

R² is H; and R³ is propargyloxy substituted phenethyl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is fluoro substituted phenethyl. In some embodiments ofa compound of Formula (I), W is

R² is H; and R³ is chloro substituted phenethyl. In some embodiments ofa compound of Formula (I), W is

R² is H; and R³ is ethynyl substituted phenethyl.

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted phenyl, substituted orunsubstituted benzyl, or substituted or unsubstituted phenethyl. In someembodiments of a compound of Formula (I), W is

R² is H; and R³ is phenyl, benzyl, or phenethyl, and R³ is substitutedwith one or more substituents selected from the group consisting ofhalo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,(C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In some embodiments of acompound of Formula (I), W is

R² is H; and R³ is phenyl or phenethyl, and R³ is substituted with onesubstituent selected from the group consisting of fluoro, chloro, bromo,methoxy, ethynyl, and propargyloxy.

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), W is

R² is H; and R³ is substituted 5-9-membered heteroaryl(C₁-C₈)alkyl. Insome embodiments of a compound of Formula (I), W is

R² is H; and R³ is unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), W is

R² is —COOR⁵ or —CONR⁵R⁶; and R³ is H. In some embodiments of a compoundof Formula (I), W is

R² is —COOR⁵; and R³ is H. In some embodiments of a compound of Formula(I), W is

R² is —CONR⁵R⁶; and R³ is H.

In some embodiments of a compound of Formula (I), W is

R² is —COOR⁵; and R⁵ is H or methyl. In some embodiments of a compoundof Formula (I), W is

R² is —COOR⁵; R⁵ is methyl; and R³ is H.

In some embodiments of a compound of Formula (I), W is

R² is —CONR⁵R⁶; R⁵ is H, methyl or ethyl; and R⁶ is phenyl.

In some embodiments of a compound of Formula (I), W is

R² is H; and R³ is H.

In some embodiments of a compound of Formula (I), W is

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl or substituted orunsubstituted 5-9-membered heteroaryl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl, or substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, orsubstituted or unsubstituted di(C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of Formula (I), W is

and R⁴ is mono-substituted (C₆-C₁₀)aryl, mono-substituted 5-9-memberedheteroaryl, mono-substituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, ormono-substituted 5-9-membered heteroaryl(C₁-C₈)alkyl. In someembodiments of Formula (I), W is

and R⁴ is di-substituted (C₆-C₁₀)aryl, di-substituted 5-9-memberedheteroaryl, di-substituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, or di-substituted5-9-membered heteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl. In some embodimentsof a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted phenyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is unsubstituted phenyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is substituted phenyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is phenyl substituted with one or more substituents selected fromhalo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,(C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In some embodiments of acompound of Formula (I), W is

and R⁴ is phenyl substituted with one or more substituents selected fromfluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy. In someembodiments of Formula (I), R⁴ is mono-substituted phenyl. In someembodiments of Formula (I), R⁴ is di-substituted phenyl. In someembodiments of a compound of Formula (I), W is

and R⁴ is methoxy substituted phenyl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is propargyloxy substituted phenyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is fluoro substituted phenyl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is chloro substituted phenyl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is ethynyl substituted phenyl.

In some embodiments of a compound of Formula (I), W is

and R⁴ is

In some embodiments of a compound of Formula (I), W is

and R⁴ is

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted 5-9-membered heteroaryl. In someembodiments of a compound of Formula (I), W is

and R⁴ is substituted 5-9-membered heteroaryl. In some embodiments of acompound of Formula (I), W is

and R⁴ is unsubstituted 5-9-membered heteroaryl.

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted benzyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is unsubstituted benzyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is substituted benzyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is benzyl substituted with one or more substituents selected fromhalo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,(C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In some embodiments of acompound of Formula (I), W is

and R⁴ is benzyl substituted with one or more substituents selected fromfluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy. In someembodiments of Formula (I), W is

and R⁴ is mono-substituted benzyl. In some embodiments of Formula (I), Wis

and R⁴ is di-substituted benzyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is methoxy substituted benzyl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is propargyloxy substituted benzyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is fluoro substituted benzyl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is chloro substituted benzyl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is ethynyl substituted benzyl.

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted phenethyl. In some embodiments ofa compound of Formula (I), W is

and R⁴ is unsubstituted phenethyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is substituted phenethyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is phenethyl substituted with one or more substituents selectedfrom halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,(C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In some embodiments of acompound of Formula (I), W is

and R⁴ is phenethyl substituted with one or more substituents selectedfrom fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy. In someembodiments of Formula (I), W is

and R⁴ is mono-substituted phenethyl. In some embodiments of Formula(I), W is

and R⁴ is di-substituted phenethyl. In some embodiments of a compound ofFormula (I), W is

and R⁴ is methoxy substituted phenethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is propargyloxy substituted phenethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is fluoro substituted phenethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is chloro substituted phenethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is ethynyl substituted phenethyl.

In some embodiments of a compound of Formula (I), R⁴ is

In some embodiments of a compound of Formula (I), R⁴ is

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted phenyl, substituted orunsubstituted benzyl, or substituted or unsubstituted phenethyl. In someembodiments of a compound of Formula (I), W is

and R⁴ is phenyl, benzyl, or phenethyl and R⁴ is substituted with one ormore substituents selected from the group consisting of halo,(C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,(C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In some embodiments of acompound of Formula (I), W is

and R⁴ is phenyl, benzyl, or phenethyl, and R⁴ is substituted with onesubstituent selected from the group consisting of fluoro, chloro, bromo,methoxy, ethynyl, and propargyloxy.

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), W is

and R⁴ is substituted 5-9-membered heteroaryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (I), W is

and R⁴ is unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted di(C₆-C₁₀)aryl(C₁-C₈)alkyl. Insome embodiments of a compound of Formula (I), W is

and R⁴ is substituted or unsubstituted diphenylmethyl. In someembodiments of a compound of Formula (I), W is

and R⁴ is unsubstituted diphenylmethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is substituted diphenylmethyl. In some embodiments of a compoundof Formula (I), W is

and R⁴ is diphenylmethyl substituted with one or more substituentsselected from halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl,(C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In someembodiments of a compound of Formula (I), W is

and R⁴ is diphenylmethyl substituted with one or more substituentsselected from fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy.In some embodiments of Formula (I), W is

and R⁴ is mono-substituted diphenylmethyl. In some embodiments ofFormula (I), W is

and R⁴ is di-substituted diphenylmethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is methoxy substituted diphenylmethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is propargyloxy substituted phenyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is fluoro substituted diphenylmethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is chloro substituted diphenylmethyl. In some embodiments of acompound of Formula (I), W is

and R⁴ is ethynyl substituted diphenylmethyl.

In some embodiments of a compound of Formula (I), W is

and R⁴ is

In some embodiments of a compound of Formula (I), W is

In some embodiments of a compound of Formula (I), Y is CH. In someembodiments of a compound of Formula (I), Y is N.

In some embodiments of a compound of Formula (I), Ring A is a5-7-membered heterocyclyl containing 0-2 NR′. In some embodiments of acompound of Formula (I), Ring A is a 5-7-membered heterocyclylcontaining 0 NR′. In some embodiments of a compound of Formula (I), RingA is a 5-7-membered heterocyclyl containing 1 NR′. In some embodimentsof a compound of Formula (I), Ring A is a 5-7-membered heterocyclylcontaining 2 NR′. In some embodiments of a compound of Formula (I), RingA is a 5-membered heterocyclyl containing 0 NR′. In some embodiments ofa compound of Formula (I), Ring A is a 5-membered heterocyclylcontaining 1 NR′. In some embodiments of a compound of Formula (I), RingA is a 5-membered heterocyclyl containing 2 NR′. In some embodiments ofa compound of Formula (I), Ring A is a 6-membered heterocyclylcontaining 0 NR′. In some embodiments of a compound of Formula (I), RingA is a 6-membered heterocyclyl containing 1 NR′. In some embodiments ofa compound of Formula (I), Ring A is a 6-membered heterocyclylcontaining 2 NR′. In some embodiments of a compound of Formula (I), RingA is a 7-membered heterocyclyl containing 0 NR′. In some embodiments ofa compound of Formula (I), Ring A is a 7-membered heterocyclylcontaining 1 NR′. In some embodiments of a compound of Formula (I), RingA is a 7-membered heterocyclyl containing 2 NR′.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted(C₆-C₁₀)arylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryloxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted5-9-membered heterocyclylcarbonyl, or substituted or unsubstituted5-9-membered heteroarylcarbonyl.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)arylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted5-9-membered heterocyclylcarbonyl, or substituted or unsubstituted5-9-membered heteroarylcarbonyl.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryloxycarbonyl, or substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₁-C₈)alkyl, substituted or unsubstituted (C₁-C₈)alkoxy, substituted orunsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl, or substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₁-C₈)alkyl, or substituted or unsubstituted (C₁-C₈)alkoxy.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₃-C₉)cycloalkyl, substituted or unsubstituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl, or substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₃-C₉)cycloalkyl, or substituted orunsubstituted (C₃-C₉)cycloalkyl(C₁-C₈)alkyl. In some embodiments of acompound of Formula (I), Ring A is a 6-membered heterocyclyl containing1 NR′; and R′ is substituted or unsubstituted 5-9-membered heterocyclyl,or substituted or unsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₆-C₁₀)aryl, substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl,substituted or unsubstituted 5-9-membered heteroaryl, or substituted orunsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (I), Ring A is a 6-membered heterocyclylcontaining 1 NR′; and R′ is substituted or unsubstituted (C₆-C₁₀)aryl,or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted5-9-membered heteroaryl, or substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), R′ is H. In someembodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is H.

In some embodiments of a compound of Formula (I), R′ is H, substitutedor unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted(C₆-C₁₀)arylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), R′ is unsubstituted (C₁-C₈)alkyl. In some embodiments of a compoundof Formula (I), R′ is substituted (C₁-C₈)alkyl. In some embodiments of acompound of Formula (I), Ring A is a 6-membered heterocyclyl containing1 NR′; and R′ is substituted or unsubstituted (C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₁-C₈)alkylcarbonyl. In some embodiments of a compound ofFormula (I), R′ is unsubstituted (C₁-C₈)alkylcarbonyl. In someembodiments of a compound of Formula (I), R′ is substituted(C₁-C₈)alkylcarbonyl. In some embodiments of a compound of Formula (I),R′ is

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₁-C₈)alkylcarbonyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl. In some embodiments of a compound ofFormula (I), R′ is substituted (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl. In someembodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₁-C₈)alkoxycarbonyl. In some embodiments of a compoundof Formula (I), R′ is unsubstituted (C₁-C₈)alkoxycarbonyl. In someembodiments of a compound of Formula (I), R′ is substituted(C₁-C₈)alkoxycarbonyl. In some embodiments of a compound of Formula (I),R′ is tert-butoxycarbonyl. In some embodiments of a compound of Formula(I), the compound is

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted(C₁-C₈)alkoxycarbonyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₆-C₁₀)arylcarbonyl. In some embodiments of a compound ofFormula (I), R′ is unsubstituted (C₆-C₁₀)arylcarbonyl. In someembodiments of a compound of Formula (I), R′ is substituted(C₆-C₁₀)arylcarbonyl. In some embodiments of a compound of Formula (I),Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₆-C₁₀)arylcarbonyl. In some embodimentsof a compound of Formula (I), Ring A is a 6-membered heterocyclylcontaining 1 NR′; and R′ is substituted or unsubstituted phenylcarbonyl.

In some embodiments of a compound of Formula (I), R′ is phenylcarbonylsubstituted with one or more substituents selected from the groupconsisting of halo, nitro, —N(R⁵)₂, (C₁-C₈)alkyl, (C₁-C₈)alkoxy,(C₆-C₁₀)aryloxy, and 5-9-membered heteroaryloxy.

In some embodiments of a compound of Formula (I), R′ is phenylcarbonylsubstituted with one or more substituents selected from the groupconsisting of fluoro, chloro, bromo, nitro, dimethylamino, methyl, andmethoxy.

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.In some embodiments of a compound of Formula (I), R′ is substituted(C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstitutedbenzyl.

In some embodiments of a compound of Formula (I), R′ is benzylsubstituted with one or more substituents selected from the groupconsisting of halo, nitro, —N(R⁵)₂, (C₁-C₈)alkyl, (C₁-C₈)alkoxy,(C₆-C₁₀)aryloxy, and 5-9-membered heteroaryloxy.

In some embodiments of a compound of Formula (I), R′ is benzylsubstituted with one or more substituents selected from the groupconsisting of fluoro, chloro, bromo, nitro, dimethylamino, methyl,methoxy, and phenoxy.

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (I), R′ is substituted or unsubstituted5-membered heterocyclyl(C₁-C₈)alkyl. In some embodiments of a compoundof Formula (I), R′ is unsubstituted 5-membered heterocyclyl(C₁-C₈)alkyl.In some embodiments of a compound of Formula (I), R′ is substituted5-membered heterocyclyl(C₁-C₈)alkyl. In some embodiments of a compoundof Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 6-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (I), R′ is unsubstituted 6-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), R′ is substituted 6-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), R′ is4-piperidylmethyl substituted with (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 7-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (I), R′ is unsubstituted 7-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), R′ is substituted 7-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 8-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (I), R′ is unsubstituted 8-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), R′ is substituted 8-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 9-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (I), R′ is unsubstituted 9-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), R′ is substituted 9-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted5-9-membered heterocyclyl(C₁-C₈)alkyl. In some embodiments of a compoundof Formula (I), Ring A is a 6-membered heterocyclyl containing 1 NR′;and R′ is substituted or unsubstituted 5-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted 6-membered heterocyclyl(C₁-C₈)alkyl. Insome embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted7-membered heterocyclyl(C₁-C₈)alkyl. In some embodiments of a compoundof Formula (I), Ring A is a 6-membered heterocyclyl containing 1 NR′;and R′ is substituted or unsubstituted 8-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(I), Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted 9-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 5-9-membered heterocyclylcarbonyl. In some embodiments ofa compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 5-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted 5-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),R′ is substituted 5-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 6-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted 6-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),R′ is substituted 6-membered heterocyclylcarbonyl. In some embodimentsof a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (I), the compound is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 7-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted 7-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),R′ is substituted 7-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 8-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted 8-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),R′ is substituted 8-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted 9-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted 9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),R′ is substituted 9-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted6-membered heterocyclylcarbonyl. In some embodiments of a compound ofFormula (I), Ring A is a 6-membered heterocyclyl containing 1 NR′; andR′ is substituted or unsubstituted 7-membered heterocyclylcarbonyl. Insome embodiments of a compound of Formula (I), Ring A is a 6-memberedheterocyclyl containing 1 NR′; and R′ is substituted or unsubstituted8-membered heterocyclylcarbonyl. In some embodiments of a compound ofFormula (I), Ring A is a 6-membered heterocyclyl containing 1 NR′; andR′ is substituted or unsubstituted 9-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (I), each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′. In someembodiments of a compound of Formula (I), each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₁-C₈)alkoxy, substituted or unsubstituted (C₁-C₈)alkylcarbonyl,substituted or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted orunsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted (C₆-C₁₀)aryl,substituted or unsubstituted (C₆-C₁₀)arylcarbonyl, substituted orunsubstituted (C₆-C₁₀)aryloxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted5-9-membered heterocyclyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl, substituted or unsubstituted 5-9-memberedheteroaryl, substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheteroarylcarbonyl.

In some embodiments of a compound of Formula (I), each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl. In some embodiments ofa compound of Formula (I), each m is 1; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl; and R⁴ is substitutedor unsubstituted (C₆-C₁₀)aryl, or substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N; Wis

and Ring A is a 6-membered heterocyclyl containing 1 NR′. In someembodiments of a compound of Formula (I), each m is 1; Y is N; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₁-C₈)alkoxy, substituted or unsubstituted (C₁-C₈)alkylcarbonyl,substituted or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted orunsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted (C₆-C₁₀)aryl,substituted or unsubstituted (C₆-C₁₀)arylcarbonyl, substituted orunsubstituted (C₆-C₁₀)aryloxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted5-9-membered heterocyclyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl, substituted or unsubstituted 5-9-memberedheteroaryl, substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheteroarylcarbonyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N; Wis

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is N; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N; Wis

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is N; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N; Wis

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl. In some embodiments ofa compound of Formula (I), each m is 1; Y is N; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl; and R⁴ is substitutedor unsubstituted (C₆-C₁₀)aryl, or substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′. In someembodiments of a compound of Formula (I), each m is 1; Y is CH; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₁-C₈)alkoxy, substituted or unsubstituted (C₁-C₈)alkylcarbonyl,substituted or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted orunsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted (C₆-C₁₀)aryl,substituted or unsubstituted (C₆-C₁₀)arylcarbonyl, substituted orunsubstituted (C₆-C₁₀)aryloxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted5-9-membered heterocyclyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl, substituted or unsubstituted 5-9-memberedheteroaryl, substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheteroarylcarbonyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is CH; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is CH; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl. In some embodiments ofa compound of Formula (I), each m is 1; Y is CH; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl; and R⁴ is substitutedor unsubstituted (C₆-C₁₀)aryl, or substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′. In someembodiments of a compound of Formula (I), each m is 1; Y is N; R¹ is H;W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₁-C₈)alkoxy, substituted or unsubstituted (C₁-C₈)alkylcarbonyl,substituted or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted orunsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted (C₆-C₁₀)aryl,substituted or unsubstituted (C₆-C₁₀)arylcarbonyl, substituted orunsubstituted (C₆-C₁₀)aryloxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted5-9-membered heterocyclyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl, substituted or unsubstituted 5-9-memberedheteroaryl, substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheteroarylcarbonyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is N; R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is N; R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is N;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl. In some embodiments ofa compound of Formula (I), each m is 1; Y is N; R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl; and R⁴ is substitutedor unsubstituted (C₆-C₁₀)aryl, or substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′. In someembodiments of a compound of Formula (I), each m is 1; Y is CH; R¹ is H;W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₁-C₈)alkoxy, substituted or unsubstituted (C₁-C₈)alkylcarbonyl,substituted or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted orunsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted (C₆-C₁₀)aryl,substituted or unsubstituted (C₆-C₁₀)arylcarbonyl, substituted orunsubstituted (C₆-C₁₀)aryloxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted5-9-membered heterocyclyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl, substituted or unsubstituted 5-9-memberedheteroaryl, substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheteroarylcarbonyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is CH; R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ is H,substituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (I),each m is 1; Y is CH; R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₁-C₈)alkyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, or substituted or unsubstituted 5-9-memberedheterocyclylcarbonyl; and R⁴ is substituted or unsubstituted(C₆-C₁₀)aryl, or substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (I), each m is 1; Y is CH;R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; and R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl. In some embodiments ofa compound of Formula (I), each m is 1; Y is CH; R¹ is H; W is

and Ring A is a 6-membered heterocyclyl containing 1 NR′; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl, or substituted orunsubstituted 5-9-membered heterocyclylcarbonyl; and R⁴ is substitutedor unsubstituted (C₆-C₁₀)aryl, or substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl.

One embodiment provides a compound of Formula (Ia):

wherein

-   -   X is halo;    -   n is 0, 1, or 2;    -   each m is independently 0, 1, or 2;    -   Y is N or CH;    -   ring A is a 5-7 membered heterocyclyl containing 0-2 additional        nitrogen atoms NR′, wherein R′ is independently at each        occurrence H, (C₁-C₈)alkyl, (C₁-C₈)alkylcarbonyl,        (C₁-C₈)alkoxycarbonyl, (C₃-C₉)cycloalkyl,        (C₃-C₉)cycloalkylcarbonyl, (C₃-C₉)cycloalkoxycarbonyl,        (C₆-C₁₀)aryl, (C₆-C₁₀)arylcarbonyl, (C₆-C₁₀)aryloxycarbonyl,        (C₆-C₁₀)aryl(C₁-C₈)alkyl, (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl,        (C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, 5-9-membered heterocyclyl,        5-9-membered heterocyclylcarbonyl, 5-9-membered heteroaryl, or        5-9-membered heteroarylcarbonyl, wherein any alkyl, cycloalkyl,        aryl, heterocyclyl, or heteroaryl can be substituted or        unsubstituted.

In some embodiments of a compound of Formula (Ia), X is chloro. In someembodiments of a compound of Formula (Ia), n=1. In some embodiments of acompound of Formula (Ia), m=1. In some embodiments of a compound ofFormula (Ia), Y is N.

In some embodiments of a compound of Formula (Ia), the compound has astructure of Formula (Ia-1)

In some embodiments of a compound of Formula (Ia-1), R′ is atert-butoxycarbonyl (Boc) group.

In some embodiments of a compound of Formula (Ia-1), the compound is:

In some embodiments of a compound of Formula (Ia-1), the compound is

One embodiment provides a compound of Formula (Ib)

wherein

-   -   each m is independently 0, 1, or 2;    -   R⁴ is substituted or unsubstituted (C₆-C₁₀)aryl, substituted or        unsubstituted 5-9-membered heteroaryl, substituted or        unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or        unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl, substituted        or unsubstituted di(C₆-C₁₀)aryl(C₁-C₈)alkyl;    -   R′ is independently H, substituted or unsubstituted        (C₁-C₈)alkyl, substituted or unsubstituted (C₁-C₈)alkoxy,        substituted or unsubstituted (C₁-C₈)alkylcarbonyl, substituted        or unsubstituted (C₁-C₈)alkoxycarbonyl, substituted or        unsubstituted (C₃-C₉)cycloalkyl, substituted or unsubstituted        (C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted or unsubstituted        (C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted        (C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted        (C₆-C₁₀)aryl, substituted or unsubstituted (C₆-C₁₀)arylcarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryloxycarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl,        substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl,        substituted or unsubstituted 5-9-membered heterocyclyl,        substituted or unsubstituted 5-9-membered        heterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted        5-9-membered heterocyclylcarbonyl, substituted or unsubstituted        5-9-membered heteroaryl, substituted or unsubstituted        5-9-membered heteroaryl(C₁-C₈)alkyl, or substituted or        unsubstituted 5-9-membered heteroarylcarbonyl.

In some embodiments of a compound of Formula (Ib), each m=1.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₁-C₈)alkyl, substituted or unsubstituted (C₁-C₈)alkoxy,substituted or unsubstituted (C₃-C₉)cycloalkyl, substituted orunsubstituted (C₃-C₉)cycloalkyl(C₁-C₈)alkyl, substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, substituted or unsubstituted5-9-membered heterocyclyl, substituted or unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl, substituted or unsubstituted 5-9-memberedheteroaryl, or substituted or unsubstituted 5-9-memberedheteroaryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is substituted or unsubstituted (C₁-C₈)alkyl, or substituted orunsubstituted (C₁-C₈)alkoxy. In some embodiments of a compound ofFormula (Ib), R′ is substituted or unsubstituted (C₃-C₉)cycloalkyl, orsubstituted or unsubstituted (C₃-C₉)cycloalkyl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heterocyclyl, or substituted or unsubstituted5-9-membered heterocyclyl(C₁-C₈)alkyl. In some embodiments of a compoundof Formula (Ib), R′ is substituted or unsubstituted (C₆-C₁₀)aryl, orsubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heteroaryl, or substituted or unsubstituted5-9-membered heteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkylcarbonyl, substituted or unsubstituted(C₃-C₉)cycloalkoxycarbonyl, substituted or unsubstituted(C₆-C₁₀)arylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryloxycarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, substituted or unsubstituted5-9-membered heterocyclylcarbonyl, or substituted or unsubstituted5-9-membered heteroarylcarbonyl. In some embodiments of a compound ofFormula (Ib), R′ is substituted or unsubstituted (C₁-C₈)alkylcarbonyl,substituted or unsubstituted (C₃-C₉)cycloalkylcarbonyl, substituted orunsubstituted (C₆-C₁₀)arylcarbonyl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, substituted or unsubstituted5-9-membered heterocyclylcarbonyl, or substituted or unsubstituted5-9-membered heteroarylcarbonyl. In some embodiments of a compound ofFormula (Ib), R′ is substituted or unsubstituted (C₁-C₈)alkoxycarbonyl,substituted or unsubstituted (C₃-C₉)cycloalkoxycarbonyl, substituted orunsubstituted (C₆-C₁₀)aryloxycarbonyl, or substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is H.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is unsubstituted (C₁-C₈)alkyl. In some embodiments of acompound of Formula (Ib), R′ is substituted (C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₁-C₈)alkoxy. In some embodiments of a compound ofFormula (Ib), R′ is substituted (C₁-C₈)alkoxy. In some embodiments of acompound of Formula (Ib), R′ is unsubstituted (C₁-C₈)alkoxy.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₁-C₈)alkylcarbonyl. In some embodiments of a compound ofFormula (Ib), R′ is unsubstituted (C₁-C₈)alkylcarbonyl. In someembodiments of a compound of Formula (Ib), R′ is substituted(C₁-C₈)alkylcarbonyl. In some embodiments of a compound of Formula (I),R′ is

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₁-C₈)alkoxycarbonyl. In some embodiments of a compoundof Formula (Ib), R′ is unsubstituted (C₁-C₈)alkoxycarbonyl. In someembodiments of a compound of Formula (I), R′ is substituted(C₁-C₈)alkoxycarbonyl. In some embodiments of a compound of Formula(Ib), R′ is tert-butoxycarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₃-C₉)cycloalkyl. In some embodiments of a compound ofFormula (Ib), R′ is substituted (C₃-C₉)cycloalkyl. In some embodimentsof a compound of Formula (Ib), R′ is unsubstituted (C₃-C₉)cycloalkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₃-C₉)cycloalkyl(C₁-C₈)alkyl. In some embodiments of acompound of Formula (Ib), R′ is substituted(C₃-C₉)cycloalkyl(C₁-C₈)alkyl. In some embodiments of a compound ofFormula (Ib), R′ is unsubstituted (C₃-C₉)cycloalkyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₃-C₉)cycloalkylcarbonyl. In some embodiments of acompound of Formula (Ib), R′ is substituted (C₃-C₉)cycloalkylcarbonyl.In some embodiments of a compound of Formula (Ib), R′ is unsubstituted(C₃-C₉)cycloalkylcarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₃-C₉)cycloalkoxycarbonyl. In some embodiments of acompound of Formula (Ib), R′ is substituted (C₃-C₉)cycloalkoxycarbonyl.In some embodiments of a compound of Formula (Ib), R′ is unsubstituted(C₃-C₉)cycloalkoxycarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)aryl. In some embodiments of a compound of Formula(Ib), R′ is substituted (C₆-C₁₀)aryl. In some embodiments of a compoundof Formula (Ib), R′ is unsubstituted (C₆-C₁₀)aryl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)arylcarbonyl. In some embodiments of a compound ofFormula (I), R′ is unsubstituted (C₆-C₁₀)arylcarbonyl. In someembodiments of a compound of Formula (I), R′ is substituted(C₆-C₁₀)arylcarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is phenylcarbonylsubstituted with one or more substituents selected from the groupconsisting of halo, nitro, —N(R⁵)₂, (C₁-C₈)alkyl, (C₁-C₈)alkoxy,(C₆-C₁₀)aryloxy, and 5-9-membered heteroaryloxy.

In some embodiments of a compound of Formula (Ib), R′ is phenylcarbonylsubstituted with one or more substituents selected from the groupconsisting of fluoro, chloro, bromo, nitro, dimethylamino, methyl, andmethoxy.

In some embodiments of a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)aryloxycarbonyl. In some embodiments of a compoundof Formula (Ib), R′ is substituted (C₆-C₁₀)aryloxycarbonyl. In someembodiments of a compound of Formula (Ib), R′ is unsubstituted(C₆-C₁₀)aryloxycarbonyl.

In some embodiments of a compound of Formula (I), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of acompound of Formula (Ib), R′ is unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl.In some embodiments of a compound of Formula (I), R′ is substituted(C₆-C₁₀)aryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is benzylsubstituted with one or more substituents selected from the groupconsisting of halo, nitro, —N(R⁵)₂, (C₁-C₈)alkyl, (C₁-C₈)alkoxy,(C₆-C₁₀)aryloxy, and 5-9-membered heteroaryloxy.

In some embodiments of a compound of Formula (Ib), R′ is benzylsubstituted with one or more substituents selected from the groupconsisting of fluoro, chloro, bromo, nitro, dimethylamino, methyl,methoxy, and phenoxy.

In some embodiments of a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted phenethyl. In some embodiments of a compound of Formula(Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl. In some embodiments of acompound of Formula (I), R′ is unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl. In some embodiments of a compound ofFormula (Ib), R′ is substituted (C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl. Insome embodiments of a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl. In some embodiments ofa compound of Formula (Ib), R′ is substituted(C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl. In some embodiments of a compound ofFormula (Ib), R′ is unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl.

In some embodiments of a compound of Formula (Ib), the compound is:

In some embodiments of a compound of Formula (Ib), the compound is:

In some embodiments of a compound of Formula (Ib), the compound is:

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (Ib), R′ is substituted or unsubstituted5-membered heterocyclyl(C₁-C₈)alkyl. In some embodiments of a compoundof Formula (Ib), R′ is unsubstituted 5-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is substituted 5-membered heterocyclyl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 6-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (Ib), R′ is unsubstituted 6-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is substituted 6-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is4-piperidylmethyl substituted with (C₆-C₁₀)aryl(C₁-C₈)alkyl. In someembodiments of a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), the compound is:

In some embodiments of a compound of Formula (Ib), the compound is:

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 7-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (Ib), R′ is unsubstituted 7-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is substituted 7-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 8-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (Ib), R′ is unsubstituted 8-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is substituted 8-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 9-membered heterocyclyl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (Ib), R′ is unsubstituted 9-memberedheterocyclyl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is substituted 9-membered heterocyclyl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (Ib), R′ is unsubstituted 5-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (Ib),R′ is substituted 5-membered heterocyclylcarbonyl. In some embodimentsof a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 6-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (Ib), R′ is unsubstituted 6-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (Ib),R′ is substituted 6-membered heterocyclylcarbonyl. In some embodimentsof a compound of Formula (I), R′ is

In some embodiments of a compound of Formula (Ib), R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 7-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (Ib), R′ is unsubstituted 7-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (Ib),R′ is substituted 7-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 8-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (Ib), R′ is unsubstituted 8-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (Ib),R′ is substituted 8-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 9-membered heterocyclylcarbonyl. In some embodiments of acompound of Formula (Ib), R′ is unsubstituted 9-memberedheterocyclylcarbonyl. In some embodiments of a compound of Formula (Ib),R′ is substituted 9-membered heterocyclylcarbonyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heteroaryl. In some embodiments of a compoundof Formula (Ib), R′ is substituted 5-9-membered heteroaryl. In someembodiments of a compound of Formula (Ib), R′ is unsubstituted5-9-membered heteroaryl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (Ib), R′ is substituted 5-9-memberedheteroaryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R′ is unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heteroarylcarbonyl. In some embodiments of acompound of Formula (Ib), R′ is substituted 5-9-memberedheteroarylcarbonyl. In some embodiments of a compound of Formula (Ib),R′ is unsubstituted 5-9-membered heteroarylcarbonyl.

In some embodiments of a compound of Formula (Ib), R⁴ is substituted orunsubstituted (C₆-C₁₀)aryl, substituted or unsubstituted(C₆-C₁₀)aryl(C₁-C₈)alkyl, or substituted or unsubstituteddi(C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R⁴ is substituted or unsubstituted 5-9-membered heteroaryl, orsubstituted or unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R⁴ is substituted orunsubstituted (C₆-C₁₀)aryl. In some embodiments of a compound of Formula(Ib), R⁴ is substituted or unsubstituted phenyl. In some embodiments ofa compound of Formula (Ib), R⁴ is unsubstituted phenyl. In someembodiments of a compound of Formula (Ib), R⁴ is substituted phenyl. Insome embodiments of a compound of Formula (Ib), R⁴ is phenyl substitutedwith one or more substituents selected from halo, (C₁-C₈)alkyl,(C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy, and(C₂-C₆)alkynyloxy. In some embodiments of a compound of Formula (Ib), R⁴is phenyl substituted with one or more substituents selected fromfluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy. In someembodiments of Formula (Ib), R⁴ is mono-substituted phenyl. In someembodiments of Formula (Ib), R⁴ is di-substituted phenyl. In someembodiments of a compound of Formula (Ib), R⁴ is methoxy substitutedphenyl. In some embodiments of a compound of Formula (Ib), R⁴ ispropargyloxy substituted phenyl. In some embodiments of a compound ofFormula (Ib), R⁴ is fluoro substituted phenyl. In some embodiments of acompound of Formula (Ib), R⁴ is chloro substituted phenyl. In someembodiments of a compound of Formula (Ib), R⁴ is ethynyl substitutedphenyl.

In some embodiments of a compound of Formula (Ib), R⁴ is

In some embodiments of a compound of Formula (Ib), R⁴ is

In some embodiments of a compound of Formula (Ib), R⁴ is substituted orunsubstituted 5-9-membered heteroaryl. In some embodiments of a compoundof Formula (Ib), R⁴ is substituted 5-9-membered heteroaryl. In someembodiments of a compound of Formula (Ib), R⁴ is unsubstituted5-9-membered heteroaryl.

In some embodiments of a compound of Formula (Ib), R⁴ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of acompound of Formula (Ib), R⁴ is substituted or unsubstituted benzyl. Insome embodiments of a compound of Formula (Ib), R⁴ is unsubstitutedbenzyl. In some embodiments of a compound of Formula (Ib), R⁴ issubstituted benzyl. In some embodiments of a compound of Formula (Ib),R⁴ is benzyl substituted with one or more substituents selected fromhalo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,(C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy. In some embodiments of acompound of Formula (Ib), R⁴ is benzyl substituted with one or moresubstituents selected from fluoro, chloro, bromo, methoxy, ethynyl, andpropargyloxy. In some embodiments of Formula (Ib), R⁴ ismono-substituted benzyl. In some embodiments of Formula (Ib), R⁴ isdi-substituted benzyl. In some embodiments of a compound of Formula(Ib), R⁴ is methoxy substituted benzyl. In some embodiments of acompound of Formula (Ib), R⁴ is propargyloxy substituted benzyl. In someembodiments of a compound of Formula (Ib), R⁴ is fluoro substitutedbenzyl. In some embodiments of a compound of Formula (Ib), R⁴ is chlorosubstituted benzyl. In some embodiments of a compound of Formula (Ib),R⁴ is ethynyl substituted benzyl.

In some embodiments of a compound of Formula (Ib), R⁴ is substituted orunsubstituted phenethyl. In some embodiments of a compound of Formula(Ib), R⁴ is unsubstituted phenethyl. In some embodiments of a compoundof Formula (Ib), R⁴ is substituted phenethyl. In some embodiments of acompound of Formula (Ib), R⁴ phenethyl substituted with one or moresubstituents selected from is halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy,(C₂-C₆)alkenyl, (C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy, and(C₂-C₆)alkynyloxy. In some embodiments of a compound of Formula (Ib), R⁴is phenethyl substituted with one or more substituents selected fromfluoro, chloro, bromo, methoxy, ethynyl, or propargyloxy. In someembodiments of Formula (Ib), R⁴ is mono-substituted phenethyl. In someembodiments of Formula (Ib), R⁴ is di-substituted phenethyl. In someembodiments of a compound of Formula (Ib), R⁴ is methoxy substitutedphenethyl. In some embodiments of a compound of Formula (Ib), R⁴ ispropargyloxy substituted phenethyl. In some embodiments of a compound ofFormula (Ib), R⁴ is fluoro substituted phenethyl. In some embodiments ofa compound of Formula (Ib), R⁴ is chloro substituted phenethyl. In someembodiments of a compound of Formula (Ib), R⁴ is ethynyl substitutedphenethyl.

In some embodiments of a compound of Formula (Ib), R⁴ is

In some embodiments of a compound of Formula (Ib), R⁴ is

In some embodiments of a compound of Formula (Ib), R⁴ is substituted orunsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl. In some embodimentsof a compound of Formula (Ib), R⁴ is substituted 5-9-memberedheteroaryl(C₁-C₈)alkyl. In some embodiments of a compound of Formula(Ib), R⁴ is unsubstituted 5-9-membered heteroaryl(C₁-C₈)alkyl.

In some embodiments of a compound of Formula (Ib), R⁴ is substituted orunsubstituted di(C₆-C₁₀)aryl(C₁-C₈)alkyl. In some embodiments of acompound of Formula (Ib), R⁴ is substituted or unsubstituteddiphenylmethyl. In some embodiments of a compound of Formula (Ib), R⁴ isunsubstituted diphenylmethyl. In some embodiments of a compound ofFormula (Ib), R⁴ is substituted diphenylmethyl. In some embodiments of acompound of Formula (Ib), R⁴ is diphenylmethyl substituted with one ormore substituents selected from halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy,(C₂-C₆)alkenyl, (C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy, and(C₂-C₆)alkynyloxy. In some embodiments of a compound of Formula (Ib), R⁴is diphenylmethyl substituted with one or more substituents selectedfrom fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy. In someembodiments of Formula (I), R⁴ is mono-substituted diphenylmethyl. Insome embodiments of Formula (Ib), R⁴ is di-substituted diphenylmethyl.In some embodiments of a compound of Formula (Ib), R⁴ is methoxysubstituted diphenylmethyl. In some embodiments of a compound of Formula(Ib), R⁴ is propargyloxy substituted phenyl. In some embodiments of acompound of Formula (Ib), R⁴ is fluoro substituted diphenylmethyl. Insome embodiments of a compound of Formula (Ib), R⁴ is chloro substituteddiphenylmethyl. In some embodiments of a compound of Formula (Ib), R⁴ isethynyl substituted diphenylmethyl.

In some embodiments of a compound of Formula (Ib), R⁴ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl and R⁴ is substituted orunsubstituted (C₆-C₁₀)aryl. In some embodiments of a compound of Formula(Ib), R′ is benzyl substituted with one or more substituents selectedfrom the group consisting of halo, (C₁-C₈)alkyl, (C₁-C₈)alkoxy,(C₆-C₁₀)aryloxy, and 5-9-membered heteroaryloxy; and R⁴ is phenylsubstituted with one substituent selected from the group consisting offluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy.

In some embodiments of a compound of Formula (Ib), R′ is benzylsubstituted with phenoxy; and R⁴ is phenyl substituted with onesubstituent selected from the group consisting of methoxy andpropargyloxy. In some embodiments of a compound of Formula (Ib), R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R⁴ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R⁴ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R′ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl; and R⁴ issubstituted or unsubstituted (C₆-C₁₀)aryl.

In some embodiments of a compound of Formula (Ib), R′ is4-piperidylmethyl substituted with (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R⁴ isphenyl substituted with one substituent selected from the groupconsisting of fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy.

In some embodiments of a compound of Formula (Ib), R′ is

and R⁴ is phenyl substituted with one substituent selected from thegroup consisting of methoxy and propargyloxy. In some embodiments of acompound of Formula (Ib), R′ is substituted or unsubstituted5-9-membered heterocyclyl(C₁-C₈)alkyl; and R⁴ is

In some embodiments of a compound of Formula (Ib), R′ is substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl; and R⁴ is

In some embodiments of a compound of Formula (Ib), each m is 1; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R⁴ issubstituted or unsubstituted (C₆-C₁₀)aryl. In some embodiments of acompound of Formula (Ib), each m is 1; R′ is benzyl substituted with oneor more substituents selected from the group consisting of halo,(C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₆-C₁₀)aryloxy, and 5-9-memberedheteroaryloxy; and R⁴ is phenyl substituted with one substituentselected from the group consisting of fluoro, chloro, bromo, methoxy,ethynyl, and propargyloxy.

In some embodiments of a compound of Formula (Ib), each m is 1; R′ isbenzyl substituted with phenoxy; and R⁴ is phenyl substituted with onesubstituent selected from the group consisting of methoxy andpropargyloxy. In some embodiments of a compound of Formula (Ib), each mis 1; R′ is substituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl; andR⁴ is

In some embodiments of a compound of Formula (Ib), each m is 1; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R⁴ is

In some embodiments of a compound of Formula (Ib), each m is 1; R′ issubstituted or unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R′ is

In some embodiments of a compound of Formula (Ib), each m is 1; R′ issubstituted or unsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl; andR⁴ is substituted or unsubstituted (C₆-C₁₀)aryl.

In some embodiments of a compound of Formula (Ib), each m is 1; R′ is4-piperidylmethyl substituted with (C₆-C₁₀)aryl(C₁-C₈)alkyl; and R⁴ isphenyl substituted with one substituent selected from the groupconsisting of fluoro, chloro, bromo, methoxy, ethynyl, and propargyloxy.

In some embodiments of a compound of Formula (Ib), each m is 1; R′ is

and R⁴ is phenyl substituted with one substituent selected from thegroup consisting of methoxy and propargyloxy. In some embodiments of acompound of Formula (Ib), each m is 1; R′ is substituted orunsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl; and R⁴ is

In some embodiments of a compound of Formula (Ib), each m is 1; R′ issubstituted or unsubstituted 5-9-membered heterocyclyl(C₁-C₈)alkyl; andR⁴ is

Further embodiments provided herein include combinations of one or moreof the particular embodiments set forth above.

In some embodiments, the compound disclosed herein has the structureprovided in Table 1.

TABLE 1 Cpd. Structure Name 1

7-(morpholine-4-carbonyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4-chloro- phenethyl)piperidine-1- carboxylate 2

1,3-dioxo-7-(piperidine-1- carbonyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- chlorophenethyl)piperidine-1- carboxylate 3

1,3-dioxo-7-(pyrrolidine-1- carbonyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- chlorophenethyl)piperidine-1- carboxylate 4

7-(3,3-dimethylbutanoyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- chlorophenethyl)piperidine-1- carboxylate 5

tert-butyl 2-(4-(4- chlorophenethyl)piperidine-1- carbonyloxy)-1,3-dioxohexahydroimidazo[1,5- a]pyrazine-7(1H)-carboxylate 6

1,3-dioxo-7-(2- phenylacetyl)hexahydroimidazo [1,5-a]pyrazin-2(3H)-yl4-(4- chlorophenethyl)piperidine-1- carboxylate 7

7-(4-bromobenzoyl)-1,3- dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl4-(4- chlorophenethyl)piperidine-1- carboxylate 8

7-(4-methylbenzoyl)-1,3- dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl4-(4- chlorophenethyl)piperidine-1- carboxylate 9

7-(4-fluoro-3-nitrobenzoyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- chlorophenethyl)piperidine-1- carboxylate 10

7-(4-(dimethylamino)benzoyl)- 1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- chlorophenethyl)piperidine-1- carboxylate 11

7-(4-methoxybenzoyl)-1,3- dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl4-(4- chlorophenethyl)piperidine-1- carboxylate 12

1,3-dioxo-7- phenethylhexahydroimidazo[1,5- a]pyrazin-2(3H)-yl 4-(4-chlorophenethyl)piperidine-1- carboxylate 13

7-(4-bromobenzyl)-1,3- dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl4-(4- chlorophenethyl)piperidine-1- carboxylate 14

tert-butyl 1,3-dioxo-2- (piperidine-1- carbonyloxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)- carboxylate 15

1-(7-(tert-butoxycarbonyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl) 4-methyl piperidine-1,4-dicarboxylate 16

7-(tert-butoxycarbonyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 3- benzylmorpholine-4-carboxylate 17

tert-butyl 1,3-dioxo-2-(4- phenethylpiperazine-1-carbonyloxy)hexahydroimidazo [1,5-a]pyrazine-7(1H)- carboxylate 18

tert-butyl 2-(4-(bis(4- chlorophenyl)methyl)piperazine-1-carbonyloxy)-1,3- dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate 19

tert-butyl 1,3-dioxo-2-(4- (phenylcarbamoyl)piperidine-1-carbonyloxy)hexahydroimidazo [1,5-a]pyrazine-7(1H)- carboxylate 20

tert-butyl 2-(4- (ethyl(phenyl)carbamoyl)piper-idine-1-carbonyloxy)-1,3- dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate 21

tert-butyl 2-((4-(4- methoxyphenyl)piperazine-1- carbonyl)oxy)-1,3-dioxohexahydroimidazo[1,5- a]pyrazine-7(1H)-carboxylate 22

1,3-dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl 4-(4-methoxyphenyl)piperazine-1- carboxylate 23

7-methyl-1,3- dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl 4-(4-methoxyphenyl)piperazine-1- carboxylate 24

7-((1-(tert- butoxycarbonyl)piperidin-4- yl)methyl)-1,3-dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl 4-(4-methoxyphenyl)piperazine-1- carboxylate 25

1,3-dioxo-7-(piperidin-4- ylmethyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- methoxyphenyl)piperidine-1- carboxylate 26

7-((1-benzylpiperidin-4- yl)methyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- methoxyphenyl)piperazine-1- carboxylate 27

tert-butyl 1,3-dioxo-2-((4- phenylpiperidine-1-carbonyl)oxy)hexahydroimidazo [1,5-a]pyrazine-7(1H)- carboxylate 28

1,3-dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl 4-phenylpiperidine-1-carboxylate 29

1-(7-(tert-Butoxycarbonyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl) 4-methyl piperidine-1,4-dicarboxylate 30

1-(1,3- Dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl) 4-methylpiperidine-1,4-dicarboxylate 31

1-(1,3-dioxo-7-(4- phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl) 4- methyl piperidine-1,4- dicarboxylate 32

7-(tert-butoxycarbonyl)-1,3- dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl morpholine- 4-carboxylate 33

1,3-dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl morpholine-4-carboxylate 34

1,3-dioxo-7-(4- phenoxybenzyl)hexahydroimidazo [1,5-a]pyrazin-2(3H)-ylmorpholine-4-carboxylate 35

1,3-dioxo-7-(3- phenoxybenzyl)hexahydroimidazo [1,5-a]pyrazin-2(3H)-yl4-(4- chlorophenethyl)piperidine-1- carboxylate 36

1,3-dioxo-7-(3- phenoxybenzyl)hexahydroimidazo [1,5-a]pyrazin-2(3H)-yl4-(4- methoxyphenyl)piperazine-1- carboxylate 37

1,3-dioxo-7-(4- phenoxybenzyl)hexahydroimidazo [1,5-a]pyrazin-2(3H)-yl4-(4- methoxyphenyl)piperazine-1- carboxylate 38

1,3-dioxo-7-(4- phenoxybenzyl)hexahydroimidazo [1,5-a]pyrazin-2(3H)-yl4- phenylpiperidine-1-carboxylate 39

1,3-dioxo-7-(4- phenoxybenzyl)hexahydroimidazo [1,5-a]pyrazin-2(3H)-yl4-(4- (prop-2-yn-1- yloxy)phenyl)piperazine-1- carboxylate 40

(S)-7-(4-bromobenzyl)-1,3- dioxohexahydroimidazo[1,5- a]pyrazin-2(3H)-yl4-(4- chlorophenethyl)piperidine-1- carboxylate 41

(S)-7-(morpholine-4-carbonyl)- 1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-chlorophenethyl)piperazine-1- carboxylate 42

(S)-7-((1-benzylpiperidin-4- yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-methoxyphenyl)piperazine-1- carboxylate 43

(R)-7-((1-benzylpiperidin-4- yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-methoxyphenyl)piperazine-1- carboxylate 44

(S)-1,3-dioxo-7-(4- phenoxybenzyl)- hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4-(prop-2- ynyloxy)phenyl)piperazine-1- carboxylate 45

(R)-1,3-dioxo-7-(4- phenoxybenzyl)- hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4-(prop-2- ynyloxy)phenyl)piperazine-1- carboxylate 46

(S)-7-((1-benzylpiperidin-4- yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-(prop-2-ynyloxy)phenyl)piperazine-1- carboxylate 47

(R)-7-((1-benzylpiperidin-4- yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-(prop-2-ynyloxy)phenyl)piperazine-1- carboxylate 48

(S)-7-(morpholine-4-carbonyl)- 1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-ethynylphenethyl)piperidine-1- carboxylate 49

(R)-7-(morpholine-4-carbonyl)- 1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-ethynylphenethyl)piperidine-1- carboxylate 50

(S)-7-(morpholine-4-carbonyl)- 1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4-chloro- phenethyl)piperidine-1- carboxylate 51

(R)-7-(morpholine-4-carbonyl)- 1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4-chloro- phenethyl)piperidine-1- carboxylate 52

7-((1-benzylpiperidin-4- yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin- 2(3H)-yl 4-(4-methoxyphenyl)piperazine-1- carboxylate 53

7-(morpholine-4-carbonyl)-1,3- dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl 4-(4- ethynylphenethyl)piperidine-1- carboxylate

Preparation of the Compounds

The compounds used in the reactions described herein are made accordingto known organic synthesis techniques, starting from commerciallyavailable chemicals and/or from compounds described in the chemicalliterature. “Commercially available chemicals” are obtained fromstandard commercial sources including Acros Organics (Geel, Belgium),Aldrich Chemical (Milwaukee, Wis., including Sigma Chemical and Fluka),Apin Chemicals Ltd. (Milton Park, UK), Ark Pharm, Inc. (Libertyville,Ill.), Avocado Research (Lancashire, U.K.), BDH Inc. (Toronto, Canada),Bionet (Cornwall, U.K.), Chemservice Inc. (West Chester, Pa.),Combi-blocks (San Diego, Calif.), Crescent Chemical Co. (Hauppauge,N.Y.), eMolecules (San Diego, Calif.), Fisher Scientific Co.(Pittsburgh, Pa.), Fisons Chemicals (Leicestershire, UK), FrontierScientific (Logan, Utah), ICN Biomedicals, Inc. (Costa Mesa, Calif.),Key Organics (Cornwall, U.K.), Lancaster Synthesis (Windham, N.H.),Matrix Scientific, (Columbia, S.C.), Maybridge Chemical Co. Ltd.(Cornwall, U.K.), Parish Chemical Co. (Orem, Utah), Pfaltz & Bauer, Inc.(Waterbury, Conn.), Polyorganix (Houston, Tex.), Pierce Chemical Co.(Rockford, Ill.), Riedel de Haen AG (Hanover, Germany), Ryan Scientific,Inc. (Mount Pleasant, S.C.), Spectrum Chemicals (Gardena, Calif.),Sundia Meditech, (Shanghai, China), TCI America (Portland, Oreg.), TransWorld Chemicals, Inc. (Rockville, Md.), and WuXi (Shanghai, China).

Suitable reference books and treatises that detail the synthesis ofreactants useful in the preparation of compounds described herein, orprovide references to articles that describe the preparation, includefor example, “Synthetic Organic Chemistry”, John Wiley & Sons, Inc., NewYork; S. R. Sandler et al., “Organic Functional Group Preparations,” 2ndEd., Academic Press, New York, 1983; H. O. House, “Modern SyntheticReactions”, 2nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L.Gilchrist, “Heterocyclic Chemistry”, 2nd Ed., John Wiley & Sons, NewYork, 1992; J. March, “Advanced Organic Chemistry: Reactions, Mechanismsand Structure”, 4th Ed., Wiley-Interscience, New York, 1992. Additionalsuitable reference books and treatises that detail the synthesis ofreactants useful in the preparation of compounds described herein, orprovide references to articles that describe the preparation, includefor example, Fuhrhop, J. and Penzlin G. “Organic Synthesis: Concepts,Methods, Starting Materials”, Second, Revised and Enlarged Edition(1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R. V. “OrganicChemistry, An Intermediate Text” (1996) Oxford University Press, ISBN0-19-509618-5; Larock, R. C. “Comprehensive Organic Transformations: AGuide to Functional Group Preparations” 2nd Edition (1999) Wiley-VCH,ISBN: 0-471-19031-4; March, J. “Advanced Organic Chemistry: Reactions,Mechanisms, and Structure” 4th Edition (1992) John Wiley & Sons, ISBN:0-471-60180-2; Otera, J. (editor) “Modern Carbonyl Chemistry” (2000)Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. “Patai's 1992 Guide to theChemistry of Functional Groups” (1992) Interscience ISBN: 0-471-93022-9;Solomons, T. W. G. “Organic Chemistry” 7th Edition (2000) John Wiley &Sons, ISBN: 0-471-19095-0; Stowell, J. C., “Intermediate OrganicChemistry” 2nd Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2;“Industrial Organic Chemicals: Starting Materials and Intermediates: AnUllmann's Encyclopedia” (1999) John Wiley & Sons, ISBN: 3-527-29645-X,in 8 volumes; “Organic Reactions” (1942-2000) John Wiley & Sons, in over55 volumes; and “Chemistry of Functional Groups” John Wiley & Sons, in73 volumes.

Specific and analogous reactants are also identified through the indicesof known chemicals prepared by the Chemical Abstract Service of theAmerican Chemical Society, which are available in most public anduniversity libraries, as well as through on-line databases (the AmericanChemical Society, Washington, D.C.,). Chemicals that are known but notcommercially available in catalogs are optionally prepared by customchemical synthesis houses, where many of the standard chemical supplyhouses (e.g., those listed above) provide custom synthesis services. Areference for the preparation and selection of pharmaceutical salts ofthe N-hydroxy bicyclic hydantoin carbamates described herein is P. H.Stahl & C. G. Wermuth “Handbook of Pharmaceutical Salts”, VerlagHelvetica Chimica Acta, Zurich, 2002.

The N-hydroxy bicyclic hydantoin carbamates were prepared by the generalsynthetic routes described below in Schemes 1-2.

A method for preparing compounds of formula G is provided in Scheme 1.Coupling of O-benzylhydroxylamine (A) with 4-Boc-piperazine-2-carboxylicacid (B) followed by cyclization afforded intermediate C. Followingbenzyl deprotection with hydrogen in the presence of a catalytic amountof Pd/C, intermediate D was coupled with a heterocyclyl carbonylchloride to afford Intermediate E. In some embodiments, the heterocyclylcarbonyl chloride is a piperidine carbonyl chloride. In someembodiments, the heterocyclyl carbonyl chloride is a piperazine carbonylchloride. In some embodiments, the heterocyclyl carbonyl chloride is amorpholine carbonyl chloride. Intermediate E was then deprotected in thepresence of an acid to afford Intermedate F which was then furtherfunctionalized to produce a compound of formula G. In some embodiments,intermediate F was reacted with a R¹⁰⁰COCl. In some embodiments, R¹⁰⁰ isheterocyclyl selected from morpholine, piperidine, or pyrrolidine. Insome embodiments, R¹⁰⁰ is alkyl. In some embodiments, R¹⁰⁰ is aryl. Insome embodiments, R¹⁰⁰ is aralkyl. In some embodiments, intermediate Fwas reacted with R¹⁰¹CHO under reductive amination conditions. In someembodiments, R¹⁰¹ is aryl. In some embodiments, R¹⁰¹ is heterocyclyl.

An alternate method for preparing compounds of formula G is provided inScheme 2. The Boc protecting group of Intermediate C was removed in thepresence of an acid, followed by reaction with R¹⁰⁰COCl or R¹⁰¹CHO asdescribed above to afford Intermediate I. Intermediate I was thendeprotected to afford intermediate J. Intermediate J was then coupledwith a heterocyclyl carbonyl chloride to afford compound s of formula Gas described in Scheme 1.

Further Forms of N-Hydroxy Bicyclic Hydantoin Carbamates DisclosedHerein

Isomers

Furthermore, in some embodiments, the compounds described herein existas geometric isomers. In some embodiments, the compounds describedherein possess one or more double bonds. The compounds presented hereininclude all cis, trans, syn, anti, entgegen (E), and zusammen (Z)isomers as well as the corresponding mixtures thereof. In somesituations, compounds exist as tautomers. The compounds described hereininclude all possible tautomers within the formulas described herein. Insome situations, the compounds described herein possess one or morechiral centers and each center exists in the R configuration, or Sconfiguration. The compounds described herein include alldiastereomeric, enantiomeric, and epimeric forms as well as thecorresponding mixtures thereof. In additional embodiments of thecompounds and methods provided herein, mixtures of enantiomers and/ordiastereoisomers, resulting from a single preparative step, combination,or interconversion are useful for the applications described herein. Insome embodiments, the compounds described herein are prepared as theirindividual stereoisomers by reacting a racemic mixture of the compoundwith an optically active resolving agent to form a pair ofdiastereoisomeric compounds, separating the diastereomers and recoveringthe optically pure enantiomers. In some embodiments, dissociablecomplexes are preferred (e.g., crystalline diastereomeric salts). Insome embodiments, the diastereomers have distinct physical properties(e.g., melting points, boiling points, solubilities, reactivity, etc.)and are separated by taking advantage of these dissimilarities. In someembodiments, the diastereomers are separated by chiral chromatography,or preferably, by separation/resolution techniques based upondifferences in solubility. In some embodiments, the optically pureenantiomer is then recovered, along with the resolving agent, by anypractical means that would not result in racemization.

Labeled Compounds

In some embodiments, the compounds described herein exist in theirisotopically-labeled forms. In some embodiments, the methods disclosedherein include methods of treating diseases by administering suchisotopically-labeled compounds. In some embodiments, the methodsdisclosed herein include methods of treating diseases by administeringsuch isotopically-labeled compounds as pharmaceutical compositions. Insome embodiments, the methods disclosed herein include methods ofinhibiting a serine hydrolase by exposing the serine hydrolase to suchisotopically-labeled compounds. Thus, in some embodiments, the compoundsdisclosed herein include isotopically-labeled compounds, which areidentical to those recited herein, but for the fact that one or moreatoms are replaced by an atom having an atomic mass or mass numberdifferent from the atomic mass or mass number usually found in nature.Examples of isotopes that are incorporated into compounds of theinvention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, sulfur, fluorine and chloride, such as ²H, ³H, ¹³C, ¹⁴C,¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl, respectively. Compoundsdescribed herein, and the pharmaceutically acceptable salts, esters,solvate, hydrates or derivatives thereof which contain theaforementioned isotopes and/or other isotopes of other atoms are withinthe scope of this invention. Certain isotopically-labeled compounds, forexample those into which radioactive isotopes such as ³H and ¹⁴C areincorporated, are useful in drug and/or substrate tissue distributionassays. Tritiated, i. e., ³H and carbon-14, i. e., ¹⁴C, isotopes areparticularly preferred for their ease of preparation and detectability.Further, substitution with heavy isotopes such as deuterium, i.e., ²H,produces certain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements. In some embodiments, the isotopically labeled compounds,pharmaceutically acceptable salt, ester, solvate, hydrate or derivativethereof is prepared by any suitable method.

In some embodiments, the compounds described herein are labeled by othermeans, including, but not limited to, the use of chromophores orfluorescent moieties, bioluminescent labels, or chemiluminescent labels.

Pharmaceutically Acceptable Salts

In some embodiments, the compounds described herein exist as theirpharmaceutically acceptable salts. In some embodiments, the methodsdisclosed herein include methods of treating diseases by administeringsuch pharmaceutically acceptable salts. In some embodiments, the methodsdisclosed herein include methods of treating diseases by administeringsuch pharmaceutically acceptable salts as pharmaceutical compositions.The invention also provides for methods of inhibiting a serine hydrolaseby exposing the serine hydrolase to such pharmaceutically acceptablesalts.

In some embodiments, the compounds described herein possess acidic orbasic groups and therefore react with any of a number of inorganic ororganic bases, and inorganic and organic acids, to form apharmaceutically acceptable salt. In some embodiments, these salts areprepared in situ during the final isolation and purification of thecompounds of the invention, or by separately reacting a purifiedcompound in its free form with a suitable acid or base, and isolatingthe salt thus formed.

Solvates

In some embodiments, the compounds described herein exist as solvates.The invention provides for methods of treating diseases by administeringsuch solvates. The invention further provides for methods of treatingdiseases by administering such solvates as pharmaceutical compositions.The invention also provides for methods of inhibiting a serine hydrolaseby exposing the serine hydrolase to such solvates.

Solvates contain either stoichiometric or non-stoichiometric amounts ofa solvent, and, in some embodiments, are formed during the process ofcrystallization with pharmaceutically acceptable solvents such as water,ethanol, and the like. Hydrates are formed when the solvent is water, oralcoholates are formed when the solvent is alcohol. Solvates of thecompounds described herein are conveniently prepared or formed duringthe processes described herein. By way of example only, hydrates of thecompounds described herein are conveniently prepared byrecrystallization from an aqueous/organic solvent mixture, using organicsolvents including, but not limited to, dioxane, tetrahydrofuran ormethanol. In addition, the compounds provided herein exist in unsolvatedas well as solvated forms. In general, the solvated forms are consideredequivalent to the unsolvated forms for the purposes of the compounds andmethods provided herein.

Pharmaceutical Compositions

In certain embodiments, the N-hydroxy bicyclic hydantoin carbamate asdescribed herein is administered as a pure chemical. In otherembodiments, the N-hydroxy bicyclic hydantoin carbamate described hereinis combined with a pharmaceutically suitable or acceptable carrier (alsoreferred to herein as a pharmaceutically suitable (or acceptable)excipient, physiologically suitable (or acceptable) excipient, orphysiologically suitable (or acceptable) carrier) selected on the basisof a chosen route of administration and standard pharmaceutical practiceas described, for example, in Remington: The Science and Practice ofPharmacy (Gennaro, 21^(st) Ed. Mack Pub. Co., Easton, Pa. (2005)).

Accordingly, provided herein is a pharmaceutical composition comprisingat least one N-hydroxy bicyclic hydantoin carbamate described herein, ora stereoisomer, pharmaceutically acceptable salt, hydrate, solvate, orN-oxide thereof, together with one or more pharmaceutically acceptablecarriers. The carrier(s) (or excipient(s)) is acceptable or suitable ifthe carrier is compatible with the other ingredients of the compositionand not deleterious to the recipient (i.e., the subject) of thecomposition.

One embodiment provides a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound of Formula (I), or apharmaceutically acceptable salt thereof.

Another embodiment provides a pharmaceutical composition consistingessentially of a pharmaceutically acceptable carrier and a compound ofFormula (I), or a pharmaceutically acceptable salt thereof.

In certain embodiments, the N-hydroxy bicyclic hydantoin carbamate asdescribed herein is substantially pure, in that it contains less thanabout 5%, or less than about 1%, or less than about 0.1%, of otherorganic small molecules, such as contaminating intermediates orby-products that are created, for example, in one or more of the stepsof a synthesis method.

These formulations include those suitable for oral, rectal, topical,buccal, parenteral (e.g., subcutaneous, intramuscular, intradermal, orintravenous) rectal, vaginal, or aerosol administration.

Methods

Disclosed herein are methods of modulating the activity of a serinehydrolase in mammalian cells. Contemplated methods, for example,comprise exposing said enzyme to a compound described herein. In someembodiments, the method comprises contacting the mammalian cells with acompound described herein. In some embodiments, the compound utilized byone or more of the foregoing methods is one of the generic, subgeneric,or specific compounds described herein, such as a compound of Formula(I). In some embodiments, the method is an in vitro method. In somemethods, the method is an in vivo method. The ability of compoundsdescribed herein to modulate or inhibit a serine hydrolase is evaluatedby procedures known in the art and/or described herein.

Another aspect of this disclosure provides methods of treating a diseaseassociated with expression or activity of a serine hydrolase in apatient. Contemplated methods include administering a pharmaceuticallyeffective amount of a compound disclosed herein or a pharmaceuticalcomposition thereof.

In some embodiments of the methods disclosed herein, the serinehydrolase is: a lysophospholipase, such as, but not limited to, LYPLA1and LYPLA2; a serine protease, including, but not limited to, trypsin,chymotrypsin, and subtilisin; an extracellular lipase, including, butnot limited to, pancreatic lipase, hepatic lipase, gastric lipase,endothelial lipase, and lipoprotein lipase; an intracellular lipase,including, but not limited to, hormone sensitive lipase,monoacylglycerol lipase, adipose triglyceride lipase, and diacylglycerollipase; a cholinesterase, including, but not limited to,acetylcholinesterase and butyrylcholinesterase; a thioesterase,including, but not limited to, fatty acid synthase (such as palmitoyl),acyl-CoA thioesterases; a phospholipase, including, but not limited to,phospholipase A2 and platelet activating factor acetylhydrolase; aprotein or glycan hydrolase, including, but not limited to, proteinphosphate methylesterase 1, acyloxyacyl hydrolase and sialic acidacetylesterase; an amidase, including, but not limited to, fatty acidamide hydrolase; or a peptidase, including, but not limited to,dipeptidyl peptidase 4, fibroblast activation protein, andprolylendopeptidase.

In some embodiments of the methods disclosed herein, the serinehydrolase is DDHD2, FAM108B1, FAM108A1, RBBP9, NCEH1, LYPLAL1, CES2,CES3, PAFAH2, PAFAH1B2, PAFAH1B3, SCPEP1, ACHE, DPP8, PRCP, PLAT,PNPLA4, PNPLA6, PNPLA8, PARL, OVCA2, DAGLB, PREP, PREPL, ESD, PPME1,CPVL, CTSA, PPT1, PPT2, PLA2G15, ABHD3, ABHD4, ABHD6, ABHD10, ABHD11,ABHD12, ABHD16A, APEH, SIAE, ACOT2, ACOT7, DPP4, DPP7, DPP9, PLA2G7,LONP1, FASN, TPP2, MGLL, LYPLA1, LYPLA2, LIPE, LIPA, or FAAH. In someembodiments, the serine hydrolase is DDHD2. In some embodiments, theserine hydrolase is FAM108B1. In some embodiments, the serine hydrolaseis FAM108A1. In some embodiments, the serine hydrolase is RBBP9. In someembodiments, the serine hydrolase is NCEH1. In some embodiments, theserine hydrolase is LYPLAL1. In some embodiments, the serine hydrolaseis CES2. In some embodiments, the serine hydrolase is CES3. In someembodiments, the serine hydrolase is PAFAH2. In some embodiments, theserine hydrolase is PAFAH1B2. In some embodiments, the serine hydrolaseis PAFAH1B3. In some embodiments, the serine hydrolase is SCPEP1. Insome embodiments, the serine hydrolase is ACHE. In some embodiments, theserine hydrolase is DPP8. In some embodiments, the serine hydrolase isPRCP. In some embodiments, the serine hydrolase is PLAT. In someembodiments, the serine hydrolase is PNPLA4. In some embodiments, theserine hydrolase is PNPLA6. In some embodiments, the serine hydrolase isPNPLA8. In some embodiments, the serine hydrolase is PARL. In someembodiments, the serine hydrolase is OVCA2. In some embodiments, theserine hydrolase is DAGLB. In some embodiments, the serine hydrolase isPREP. In some embodiments, the serine hydrolase is PREPL. In someembodiments, the serine hydrolase is ESD. In some embodiments, theserine hydrolase is PPME1. In some embodiments, the serine hydrolase isCPVL. In some embodiments, the serine hydrolase is CTSA. In someembodiments, the serine hydrolase is PPT1. In some embodiments, theserine hydrolase is PPT2. In some embodiments, the serine hydrolase isPLA2G15. In some embodiments, the serine hydrolase is ABHD3. In someembodiments, the serine hydrolase is ABHD4. In some embodiments, theserine hydrolase is ABHD6. In some embodiments, the serine hydrolase isABHD10. In some embodiments, the serine hydrolase is ABHD11. In someembodiments, the serine hydrolase is ABHD12. In some embodiments, theserine hydrolase is ABHD16A. In some embodiments, the serine hydrolaseis APEH. In some embodiments, the serine hydrolase is SIAE. In someembodiments, the serine hydrolase is ACOT2. In some embodiments, theserine hydrolase is ACOT7. In some embodiments, the serine hydrolase isDPP4. In some embodiments, the serine hydrolase is DPP7. In someembodiments, the serine hydrolase is DPP9. In some embodiments, theserine hydrolase is PLA2G7. In some embodiments, the serine hydrolase isLONP1. In some embodiments, the serine hydrolase is FASN. In someembodiments, the serine hydrolase is TPP2. In some embodiments, theserine hydrolase is MGLL. In some embodiments, the serine hydrolase isLYPLA1. In some embodiments, the serine hydrolase is LYPLA2. In someembodiments, the serine hydrolase is LIPE. In some embodiments, theserine hydrolase is LIPA. In some embodiments, the serine hydrolase isFAAH.

Another embodiment provides a method of modulation of aprotein-palmitoyl thioesterase, comprising contacting theprotein-palmitoyl thioesterase with an effective amount or concentrationof an N-hydroxy bicyclic hydantoin carbamate described herein. In someembodiments, the protein-palmitoyl thioesterase is LYPLA1, LYPLA2, orboth. In some embodiments, the protein-palmitoyl thioesterase is PPT1.

Another embodiment provides a method of treatment of a medical conditionin a patient, wherein modulation of a protein-palmitoyl thioesterase ismedically indicated, comprising administering an effective dose of anN-hydroxy bicyclic hydantoin carbamate described herein or apharmaceutical composition thereof. In some embodiments, theprotein-palmitoyl thioesterase is LYPLA1, LYPLA2, or both. In someembodiments, the protein-palmitoyl thioesterase is PPT1.

Another embodiment provides a method of modulation of a phospholipase,comprising contacting the phospholipase with an effective amount orconcentration of an N-hydroxy bicyclic hydantoin carbamate describedherein. In some embodiments, the phospholipase is ABHD3, ABHD4, ABHD6,ABHD10, ABHD11, ABHD12, or ABHD16A. In some embodiments thephospholipase is ABHD3, ABHD4, or both.

Another embodiment provides a method of treatment of a medical conditionin a patient, wherein modulation of a phospholipase is medicallyindicated, comprising administering an effective dose of an N-hydroxybicyclic hydantoin carbamate described herein or a pharmaceuticalcomposition thereof. In some embodiments, the phospholipase is ABHD3,ABHD4, ABHD6, ABHD10, ABHD11, ABHD12, or ABHD16A. In some embodimentsthe phospholipase is ABHD3, ABHD4, or both.

Also disclosed herein are methods of treating and/or preventing in apatient in need thereof a disorder such as one or more of cancer, pain,diabetes, obesity/metabolic syndrome, epilepsy, traumatic brain injury,and inflammation. In some embodiments, the disorder is cancer.

In some embodiments, the disorder is pain. In some embodiments, thedisorder is diabetes. In some embodiments, the disorder isobesity/metabolic syndrome. In some embodiments, the disorder isepilepsy. In some embodiments, the disorder is traumatic brain injury.In some embodiments, the disorder is inflammation. In some embodiments,the disorder is pain and inflammation. In some embodiments, the disorderis pain or inflammation. Disclosed methods include administering apharmaceutically effective amount of an N-hydroxy bicyclic hydantoincarbamate described herein or a pharmaceutical composition thereof.

In certain embodiments, a disclosed compound utilized by one or more ofthe foregoing methods is one of the generic, subgeneric, or specificcompounds described herein, such as a compound of Formula (I).

Disclosed compounds are administered to patients (animals and humans) inneed of such treatment in dosages that will provide optimalpharmaceutical efficacy. It will be appreciated that the dose requiredfor use in any particular application will vary from patient to patient,not only with the particular compound or composition selected, but alsowith the route of administration, the nature of the condition beingtreated, the age and condition of the patient, concurrent medication orspecial diets then being followed by the patient, and other factors,with the appropriate dosage ultimately being at the discretion of theattendant physician. For treating clinical conditions and diseases notedabove, a contemplated compound disclosed herein is administered orally,subcutaneously, topically, parenterally, by inhalation spray or rectallyin dosage unit formulations containing conventional non-toxicpharmaceutically acceptable carriers, adjuvants and vehicles. Parenteraladministration include subcutaneous injections, intravenous orintramuscular injections or infusion techniques.

Also contemplated herein are combination therapies, for example,co-administering a disclosed compound and an additional active agent, aspart of a specific treatment regimen intended to provide the beneficialeffect from the co-action of these therapeutic agents. The beneficialeffect of the combination includes, but is not limited to,pharmacokinetic or pharmacodynamic co-action resulting from thecombination of therapeutic agents. Administration of these therapeuticagents in combination typically is carried out over a defined timeperiod (usually weeks, months or years depending upon the combinationselected). Combination therapy is intended to embrace administration ofmultiple therapeutic agents in a sequential manner, that is, whereineach therapeutic agent is administered at a different time, as well asadministration of these therapeutic agents, or at least two of thetherapeutic agents, in a substantially simultaneous manner.

Substantially simultaneous administration is accomplished, for example,by administering to the subject a single formulation or composition,(e.g., a tablet or capsule having a fixed ratio of each therapeuticagent or in multiple, single formulations (e.g., capsules) for each ofthe therapeutic agents. Sequential or substantially simultaneousadministration of each therapeutic agent is effected by any appropriateroute including, but not limited to, oral routes, intravenous routes,intramuscular routes, and direct absorption through mucous membranetissues. The therapeutic agents are administered by the same route or bydifferent routes. For example, a first therapeutic agent of thecombination selected is administered by intravenous injection while theother therapeutic agents of the combination are administered orally.Alternatively, for example, all therapeutic agents are administeredorally or all therapeutic agents are administered by intravenousinjection.

Combination therapy also embraces the administration of the therapeuticagents as described above in further combination with other biologicallyactive ingredients and non-drug therapies. Where the combination therapyfurther comprises a non-drug treatment, the non-drug treatment isconducted at any suitable time so long as a beneficial effect from theco-action of the combination of the therapeutic agents and non-drugtreatment is achieved. For example, in appropriate cases, the beneficialeffect is still achieved when the non-drug treatment is temporallyremoved from the administration of the therapeutic agents, perhaps bydays or even weeks.

The components of the combination are administered to a patientsimultaneously or sequentially. It will be appreciated that thecomponents are present in the same pharmaceutically acceptable carrierand, therefore, are administered simultaneously. Alternatively, theactive ingredients are present in separate pharmaceutical carriers, suchas, conventional oral dosage forms, that are administered eithersimultaneously or sequentially.

For example, e.g., for contemplated treatment of pain and/orinflammation, a disclosed compound is co-administered with anothertherapeutic for pain such as an opioid, a cannabinoid receptor (CB-1 orCB-2) modulator, a COX-2 inhibitor, acetaminophen, and/or anon-steroidal anti-inflammatory agent. Additional therapeutics, e.g.,for the treatment of pain that are co-administered include, but are notlimited to, morphine, codeine, hydromorphone, hydrocodone, oxymorphone,fentanyl, tramadol, and levorphanol.

Other contemplated therapeutics for co-administration include, but arenot limited to, aspirin, naproxen, ibuprofen, salsalate, diflunisal,dexibuprofen, fenoprofen, ketoprofen, oxaprozin, loxoprofen,indomethacin, tolmetin, sulindac, etodolac, ketorolac, piroxicam,meloxicam, tenoxicam, droxicam, lornoxicam, celecoxib, parecoxib,rimonabant, and/or etoricoxic.

The following examples are provided merely as illustrative of variousembodiments and shall not be construed to limit the invention in anyway.

EXAMPLES

I. Chemical Synthesis

Unless otherwise noted, all reagents, including dry solvents, werepurchased from Sigma-Aldrich, Acros, Fisher, Fluka, Thermo, or CellGroand used without further purification. All chemical synthesis reactionswere carried out under a nitrogen atmosphere using oven-dried glassware.Flash chromatography was performed using 230-400 mesh silica gel. NMRspectra were recorded on a Varian Inova-400 spectrometer and werereferenced to trimethylsilane (TMS) or the residual solvent peak.Chemical shifts are reported in ppm relative to TMS and J values arereported in Hz. High resolution mass spectrometry (HRMS) experimentswere performed on an Agilent mass spectrometer using electrosprayionization-time of flight (ESI-TOF). Mass spectrometry (MS) experimentswere performed on an Agilent 1100 series MSD. All other protocols aresummarized below.

Compound 1 was erroneously described as being commercially availablefrom Sigma as ALD00010 in U.S. Provisional Application No. 62/001,869filed on May 22, 2014 from which this international application claimspriority. Compound 1 was in fact not commercially available at the timeof filing of U.S. Provisional Application No. 62/001,869.

Example 1: Preparation of7-(morpholine-4-carbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chloro-phenethyl)piperidine-1-carboxylate (Compound 1, JJH-254)

Step 1: tert-Butyl2-(benzyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

To a stirred solution of O-benzylhydroxylamine (1.73 g, 10.8 mmol, 1equiv) and N-methylmorpholine (NMM; 1.3 mL, 1.1 equiv) in CH₂Cl₂ (55 mL)at 0° C. was added 1,1′-carbonyldiimidazole (CDI; 1.76 g, 1 equiv). Theresulting solution was allowed to warm to 25° C. and stirred for 2 h. Atthis time, 4-Boc-piperazine-2-carboxylic acid (2.0 g, 8.7 mmol, 0.8equiv) was added in one portion, followed by additional NMM (1.9 mL, 1.6equiv). The resulting suspension was stirred for 24 h at 2° C., at whichtime moisture was notably generated in the reaction vessel; Na₂SO₄ wasadded, followed by additional CH₂Cl₂ (110 mL), and the mixture wasstirred for an additional 24 h. The suspension was then filtered,concentrated by rotary evaporation under reduced pressure, and theresulting residue separated by silica gel flash chromatography (40%EtOAc/hexanes) to yield tert-butyl2-(benzyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(2.1 g, 66%) as a white solid. ¹H NMR (CDCl₃, 400 MHz) δ 7.50 (m, 2H),7.39 (m, 3H), 5.17 (s, 2H), 4.43 (bs, 1H), 4.14 (bs, 1H), 4.01 (dd,J=13.6 Hz, 3.6 Hz, 1H), 3.86 (m, 1H), 2.96 (m, 1H), 2.64 (m, 1H), 2.47(m, 1H), 1.46 (s, 9H). MS calculated for C₁₈H₂₃N₃O₅ [M+H]⁺ 362.2, found362.3.

Step 2: tert-Butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

To tert-butyl2-(benzyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(300 mg, 0.83 mmol, 1 equiv) dissolved in EtOAc (4 mL) was addedpalladium on carbon (10% w/w; 85 mg, ˜10 mol %). The suspension wassparged with hydrogen gas, and then stirred under an atmosphere ofhydrogen at ambient pressure overnight at 25° C. At this time, themixture was filtered over celite and concentrated, quantitativelyyielding tert-butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate asa crystalline white solid that was used without further purification. ¹HNMR (CDCl₃, 400 MHz) δ 4.56 (bs, 1H), 4.13 (m, 2H), 4.04 (m, 2H), 3.06(m, 1H), 2.78 (m, 2H), 1.46 (s, 9H). MS calculated for C₁₁H₁₇N₃O₅ [M+H]⁺272.1, found 272.1.

Step 3: tert-Butyl2-(4-(4-chlorophenethyl)piperidine-1-carbonyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

To a stirred solution of tert-butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(225 mg, 0.83 mmol, 1 equiv) in THF (3 mL) at 25° C. was addedtriethylamine (260 μL, ˜3 equiv), and a catalytic amount of4-dimethylaminopyridine (4-DMAP). After solubilization of the reagents,4-(4-chlorophenethyl)piperidine-1-carbonyl chloride (350 mg, 1.5 equiv)was added to the reaction, and the solution warmed to 60° C. and stirredfor 2 h. The reaction was cooled to 25° C., and the precipitate(triethylammonium hydrochloride) filtered off and washed with additionalTHF. The combined filtrates were concentrated by rotary evaporationunder reduced pressure, and the residue was used in the next stepwithout further purification. An analytical sample was purified byprep-TLC (50% EtOAc/hexanes) to yield tert-butyl2-(4-(4-chlorophenethyl)piperidine-1-carbonyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylateas a white solid. MS calculated for C₂₅H₃₃ClN₄O₆ [M+Na]⁺ 543.2, found543.2.

Step 4: 1,3-Dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate hydrochloride

tert-Butyl2-(4-(4-chlorophenethyl)piperidine-1-carbonyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylatewas dissolved in 2 N methanolic HCl (10 mL) at 0° C. The stirredsolution was allowed to warm to 25° C. over the course of 30 minutes.The solvent was removed, and the resulting1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate hydrochloride was usedwithout further purification (325 mg, 86% over 2 steps). An analyticalsample was prepared by recrystallization of the salt from MeOH/Et₂O,yielding white needles. ¹H NMR (CDCl₃, 400 MHz) δ 7.25 (d, J=8 Hz, 2H),7.09 (d, J=8 Hz, 2H), 4.85 (d, J=10.8 Hz, 1H), 4.28 (d, J=14.4 Hz, 1H),4.18 (d, J=13.6 Hz, 1H), 4.05 (m, 2H), 3.68 (m, 2H), 3.02 (m, 3H), 2.63(m, 1H) 1.45-1.81 (m, 9H), 1.33 (m, 1H). MS calculated for C₂₀H₂₅ClN₄O₄[M+H]⁺ 421.2, found 421.1.

Step 5:7-(Morpholine-4-carbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chloro-phenethyl)piperidine-1-carboxylate

1,3-Dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate hydrochloride (40 mg, 0.09mmol, 1 equiv) was suspended in THF (500 μL) at 25° C., andtriethylamine (30 μL, 3 equiv) was added, followed by a catalytic amountof 4-DMAP, and then 4-morpholinecarbonyl chloride (16 μL, 1.5 equiv).The resulting solution was heated at 60° C. for 2 h. After cooling to25° C., the triethylammonium hydrochloride was removed by filtration,and the filtrates concentrated by rotary evaporation under reducedpressure. The residue was separated by prep-TLC (100% EtOAc) to yield7-(morpholine-4-carbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chloro-phenethyl)piperidine-1-carboxylateas a white solid (39.8 mg, 85%). ¹H NMR (CDCl₃, 400 MHz) δ 7.25 (d,J=8.4 Hz, 2H), 7.09 (d, J=8.4 Hz, 2H), 4.22 (m, 2H), 4.09 (m, 2H) (4.06,s, 1H), 3.70 (m, 4H), 3.68 (s, 1H), 3.32 (m, 4H), 3.12 (m, 1H), 2.95 (m,2H), 2.87 (m, 1H), 2.61 (m, 2H), 1.79 (d, J=13.2 Hz, 2H), 1.25-1.59 (m,6H). HRMS calculated for C₂₅H₃₂ClN₅O₆ [M+H]⁺ 534.2114, found 534.2116.

Example 2: Preparation of1,3-dioxo-7-(piperidine-1-carbonyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 2, JJH-250)

The title compound was prepared by the method of Example 1 using1-piperidinecarbonyl chloride in place of 4-morpholinecarbonyl chloride.MS calculated for C₂₆H₃₄ClN₅O₅ [M+H]⁺ 532. found 532.

Example 3: Preparation of1,3-dioxo-7-(pyrrolidine-1-carbonyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 3, JJH-247)

The title compound was prepared by the method of Example 1 using1-pyrrolidinecarbonyl chloride in place of 4-morpholinecarbonylchloride. MS calculated for C₂₅H₃₂ClN₅O₅ [M+H]⁺ 518, found 518.

Example 4: Preparation of7-(3,3-dimethylbutanoyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 4, JJH-246)

The title compound was prepared by the method of Example 1 usingt-butylacetyl chloride in place of 4-morpholinecarbonyl chloride. MScalculated for C₂₆H₃₅ClN₄O₅ [M+H]⁺ 519, found 519.

Example 5: Preparation of tert-butyl2-(4-(4-chlorophenethyl)piperidine-1-carbonyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 5, JJH-221)

The title compound 5 was prepared as shown in Step 3 of Example 1. MScalculated for C₂₅H₃₃ClN₄O₆ [M+H]⁺ 521, found 521.

Example 6: Preparation of1,3-dioxo-7-(2-phenylacetyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 6, JJH-249)

The title compound was prepared by the method of Example 1 usingphenylacetyl chloride in place of 4-morpholinecarbonyl chloride. MScalculated for C₂₈H₃₁ClN₄O₅ [M+H]⁺ 539, found 539.

Example 7: Preparation of7-(4-bromobenzoyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 7, JJH-248)

The title compound was prepared by the method of Example 1 using4-bromobenzoyl chloride in place of 4-morpholinecarbonyl chloride. MScalculated for C₂₇H₂₈BrClN₄O₅ [M+H]⁺ 603, found 603.

Example 8: Preparation of7-(4-methylbenzoyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 8, JJH-258)

The title compound was prepared by the method of Example 1 using toluoylchloride in place of 4-morpholinecarbonyl chloride. MS calculated forC₂₈H₃₁ClN₄O₅ [M+H]⁺ 539, found 539.

Example 9: Preparation of7-(4-fluoro-3-nitrobenzoyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 9, JJH-259)

The title compound was prepared by the method of Example 1 using4-fluoro-3-nitrobenzoyl chloride in place of 4-morpholinecarbonylchloride. MS calculated for C₂₇H₂₇ClFN₅O₇ [M+NH₄]⁺ 605, found 605.

Example 10: Preparation of7-(4-(dimethylamino)benzoyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate(Compound 10, JJH-260)

The title compound was prepared by the method of Example 1 using4-dimethylaminobenzoyl chloride in place of 4-morpholinecarbonylchloride. MS calculated for C₂₉H₃₄ClN₅O₅ [M+H]⁺ 568, found 568.

Example 11: Preparation of7-(4-methoxybenzoyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 11, JJH-261)

The title compound was prepared by the method of Example 1 using4-methoxybenzoyl chloride in place of 4-morpholinecarbonyl chloride. MScalculated for C₂₈H₃₁ClN₄O₆ [M+H]⁺ 555, found 555.

Example 12: Preparation of1,3-dioxo-7-phenethylhexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 12, JJH-252)

The title compound was prepared by the method of Example 1, but thefinal acylation step was substituted with a reductive amination(according to the method of Example 13, see below) using1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate hydrochloride (1 equiv)(Example 1, step 4), phenylacetaldehyde (2.4 equiv), and NaBH(OAc)₃ (1.4equiv). MS calculated for C₂₈H₃₃ClN₄O₄ [M+H]⁺ 525, found 525.

Example 13: Preparation of7-(4-bromobenzyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 13, JJH-251)

The title compound was prepared by the method of Example 1, but thefinal acylation step was substituted with the following reductiveamination: 7.3 mg (0.016 mmol, 1 equiv) of1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate hydrochloride (Example 1,step 4) was suspended in 0.2 ml THF, followed by 1 μl HOAc (˜1 equiv), 7mg (2.4 equiv) 4-bromobenzaldehyde and 5.5 mg (1.4 equiv) NaBH(OAc)₃.The resultant slurry was stirred at room temperature overnight, dilutedwith sat. aq. NaHCO₃, and extracted twice with CH₂Cl₂. The combinedextracts were combined, dried over Na₂SO₄, and concentrated in vacuo.The resulting residue was purified by prep-TLC (40% EtOAc/hexanes+1%triethylamine) yielding a white solid (5.4 mg, 57%). MS calculated forC₂₇H₃₀BrClN₄O₄ [M+H]⁺ 589, found 589.

Example 14: Preparation of tert-butyl1,3-dioxo-2-(piperidine-1-carbonyloxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 14, JJH-235)

The title compound was prepared by the method of Example 1, Step 3,using 1-piperidinecarbonyl chloride in place of4-(4-chlorophenethyl)piperidine-1-carbonyl chloride. MS calculated forC₁₇H₂₆N₄O₆ [M+NH₄]⁺ 400, found 400.

Example 15: Preparation of1-(7-(tert-butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)4-methyl piperidine-1,4-dicarboxylate (Compound 15, JJH-238)

The title compound was prepared by the method Example 1, Step 3, using4-carboxymethylpiperidine carbonyl chloride in place of4-(4-chlorophenethyl)piperidine-1-carbonyl chloride. MS calculated forC₁₉H₂₈N₄O₈ [M+NH₄]⁺ 458, found 458.

Example 16: Preparation of7-(tert-butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl3-benzylmorpholine-4-carboxylate (Compound 16, JJH-240)

The title compound was prepared by the method of Example 1, Step 3,using racemic 3-benzylmorpholinecarbonyl chloride in place of4-(4-chlorophenethyl)piperidine-1-carbonyl chloride. MS calculated forC₂₃H₃₀N₄O₇ [M+NH₄]⁺ 492, found 492.

Example 17: Preparation of tert-butyl1,3-dioxo-2-(4-phenethylpiperazine-1-carbonyloxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 17, JJH-241)

The title compound was prepared by the method of Example 1, Step 3,using 4-phenethyl-1-piperazinecarbonyl chloride in place of4-(4-chlorophenethyl)piperidine-1-carbonyl chloride. MS calculated forC₂₄H₃₃N₅O₆ [M+H]⁺ 488, found 488.

Example 18: Preparation of tert-butyl2-(4-(bis(4-chlorophenyl)methyl)piperazine-1-carbonyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 18, JJH-242)

The title compound was prepared by the method of Example 1, Step 3,using 4-(bis(4-chlorophenyl)methyl)piperazine-1-carbonyl chloride inplace of 4-(4-chlorophenethyl)piperidine-1-carbonyl chloride. MScalculated for C₂₉H₃₃Cl₂N₅O₆ [M+H]⁺ 618, found 618.

Example 19: Preparation of tert-butyl1,3-dioxo-2-(4-(phenylcarbamoyl)piperidine-1-carbonyloxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 19, JJH-243)

The title compound was prepared by the method of Example 1, Step 3,using 4-(phenylcarbamoyl)piperidine-1-carbonyl chloride in place of4-(4-chlorophenethyl)piperidine-1-carbonyl chloride. MS calculated forC₂₄H₃₁N₅O₇ [M+NH₄]⁺ 519, found 519.

Example 20: Preparation of tert-butyl2-(4-(ethyl(phenyl)carbamoyl)piperidine-1-carbonyloxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 20, JJH-244)

The title compound was prepared by the method of Example 1, Step 3,using 4-(ethyl(phenyl)carbamoyl)piperidine-1-carbonyl chloride in placeof 4-(4-chlorophenethyl)piperidine-1-carbonyl chloride. MS calculatedfor C₂₆H₃₅N₅O₇ [M+NH₄]⁺ 547, found 547.

Example 21: Preparation of tert-butyl2-((4-(4-methoxyphenyl)piperazine-1-carbonyl)oxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 21, MJN202)

Step 1: 4-(4-methoxyphenyl)piperazine-1-carbonyl chloride

1-(4-Methoxyphenyl)piperazine (14.25 g, 74.1 mmol, 1 equiv) wasdissolved in 120 ml dry THF, and pyridine (30 ml, ˜5 equiv) was added,and the resulting solution was cooled to 0° C. At this time, triphosgene(11 g, 0.5 equiv) was added in small portions. The resulting mixture wasallowed to warm to room temperature and was stirred overnight. Themixture was filtered, concentrated, and the residue was separated byflash chromatography over SiO₂ to yield4-(4-methoxyphenyl)piperazine-1-carbonyl chloride (1.0 g, 5%) as a paleyellow solid. ¹H NMR (CDCl₃, 400 MHz) δ 6.88 (m, 4H), 3.90 (m, 2H), 3.81(m, 2H), 3.78 (s, 3H), 3.11 (m, 4H). HRMS calculated for C₁₂H₁₆ClN₂O₂[M+H]⁺ 255.0895, found 255.0895.

Step 2: tert-butyl2-((4-(4-methoxyphenyl)piperazine-1-carbonyl)oxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

tert-Butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Example 1, Step 2, 200 mg, 0.737 mmol, 1 equiv) was dissolved in 3.5 mldry THF, and triethylamine (225 μl, 3 equiv) and a catalytic amount of4-DMAP, followed by 4-(4-methoxyphenyl)piperazine-1-carbonyl chloride(188 mg, 1 equiv) at room temperature. The solution was heated at 60° C.for 2 h, filtered, and concentrated to a residue without furtherpurification (267 mg, 93% over two steps). A sample was separated byprep-TLC (60% EtOAc/hexanes) for analysis to yield an off-white solid.¹H NMR (CDCl3, 500 MHz) δ 6.90 (m, 2H), 6.84 (m, 2H), 4.55 (m, 1H), 4.18(m, 1H), 4.08 (m, 2H), 3.80 (m, 2H), 3.77 (s, 3H), 3.67 (m, 2H), 3.07(m, 4H), 3.02 (m, 1H), 2.84 (m, 2H), 1.46 (s, 9H). HRMS calculated forC₂₃H₃₂N₅O₇ [M+H]⁺ 490.2296, found 490.2296.

Example 22: Preparation of1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 22, JJH331)

tert-Butyl2-((4-(4-methoxyphenyl)piperazine-1-carbonyl)oxy)-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylatewas Boc deprotected using methanolic HCl as described for Example 1,Step 4, yielding a white solid, after recrystallization from MeOH/ether(266.6 mg, 93%). ¹H NMR (D₂O, 400 MHz) δ 7.47 (d, J=6.8 Hz, 2H), 7.14(d, J=6.8 Hz, 2H), 4.85 (m, 1H), 4.37 (m, 1H), 4.09 (m, 2H), 3.98 (m,1H), 3.94 (m, 2H), 3.88 (s, 3H), 3.68 (m, 1H), 3.66 (m, 4H), 3.56 (m,1H), 3.41 (m, 1H), 3.29 (m, 1H). HRMS calculated for C₁₈H₂₄N₅O₅ [M+H]⁺390.1772, found 390.1772.

Example 23: Preparation of7-methyl-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 23, MJN200)

1,3-Dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (hydrochloride; 20 mg, 0.047mmol, 1 equiv) was suspended in 0.2 ml dry THF, and formaldehyde (6 μl,37% wt. in water, 1.5 equiv), acetic acid (2.8 μl, 1 equiv) andNaBH(OAc)₃ (11 mg, 1.1 equiv) were added at room temperature. Theresulting suspension was stirred at room temperature overnight,concentrated to a residue, and separated by prep-TLC (EtOAc+1%triethylamine) to yield7-methyl-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate as an off-white solid (12mg, 63%). ¹H NMR (CDCl₃, 400 MHz) δ 6.90 (m, 2H), 6.85 (m, 2H), 4.18 (m,1H), 4.06 (m, 1H), 3.80 (m, 2H), 3.77 (s, 3H), 3.67 (m, 2H), 3.23 (m,1H), 3.17 (m, 1H), 3.14 (m, 4H), 3.06 (m, 1H), 2.80 (m, 1H), 2.37 (s,3H), 2.08 (m, 2H). HRMS calculated for C₁₉H₂₆N₅O₅ [M+H]⁺ 404.1928, found404.1930.

Example 24: Preparation of7-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 24, ABC37)

The title compound was prepared as described in Example 23, usingN-Boc-4-piperidine carboxaldehyde in place of formaldehyde. (36.4 mg,66%). ¹H NMR (CDCl₃, 500 MHz) δ 6.90 (d, J=10 Hz, 2H), 6.85 (d, J=10Hz), 4.15 (m, 2H), 4.04 (m, 2H), 3.77 (s, 3H), 3.67 (m, 2H), 3.26 (m,1H), 3.14 (m, 1H), 3.11 (m, 4H), 2.80 (m, 1H), 2.69 (m, 2H), 2.27 (m,2H), 2.12 (m, 2H), 1.68 (m, 5H), 1.46 (s, 9H), 1.10 (m, 2H). HRMScalculated for C₂₉H₄₃N₆O₇ [M+H]⁺ 587.3188, found 587.3188.

Example 25: Preparation of1,3-dioxo-7-(piperidin-4-ylmethyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperidine-1-carboxylate (Compound 25, ABC38)

7-((1-(tert-Butoxycarbonyl)piperidin-4-yl)methyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylatewas Boc-deprotected using methanolic HCl as described for Example 1,Step 4, yielding a yellow solid (100%). ¹H NMR (CD₃OD, 500 MHz) δ 7.80(m, 2H), 7.22 (m, 2H), 5.15 (m, 1H), 4.41 (m, 1H), 4.28 (m, 2H), 4.19(m, 3H), 3.94 (s, 3H), 3.90 (m, 4H), 3.86 (m, 2H), 3.54 (m, 2H), 3.38(m, 3H), 3.14 (m, 2H), 2.48 (m, 1H), 2.33 (m, 2H), 1.66 (m, 2H). HRMScalculated for C₂₄H₃₅N₆O₅ [M+H]⁺ 487.2663, found 487.2664.

Example 26: Preparation of7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 26, ABC44)

The title compound was prepared as described in Example 23, usingN-benzyl-4-piperidine carboxaldehyde in place of formaldehyde. (97 mg,34%). ¹H NMR (CDCl₃, 400 MHz) δ 7.40 (m, 5H), 6.98 (m, 2H), 6.92 (m,2H), 4.22 (m, 1H), 4.10 (m, 1H), 3.88 (m, 2H), 3.85 (s, 3H), 3.75 (m,2H), 3.62 (s, 2H), 3.33 (m, 1H), 3.19 (m, 1H), 3.18 (m, 4H), 2.99 (m,2H), 2.88 (m, 1H), 2.34 (m, 2H), 2.19 (m, 2H), 2.07 (m, 2H), 1.80 (m,2H), 1.58 (m, 1H), 1.37 (m, 2H)□ HRMS calculated for C₃₁H₄₁N₆O₅ [M+H]⁺577.3133, found 577.3133.

Example 27: Preparation of tert-butyl1,3-dioxo-2-((4-phenylpiperidine-1-carbonyl)oxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Compound 27, JJH321)

Step 1: (1H-imidazol-1-yl)(4-phenylpiperidin-1-yl)methanone

4-Phenylpiperidine (50 mg, 0.31 mmol, 1 equiv) was dissolved in 1.56 mlTHF, and triethylamine (63 ml, 2 equiv) and CDI (60 mg, 1.2 equiv) addedin order at room temperature. The resulting solution was stirred at roomtemperature overnight, and concentrated to a residue that was separatedby prep-TLC (60% acetone/hexanes) to yield(1H-imidazol-1-yl)(4-phenylpiperidin-1-yl)methanone as a crystallinewhite solid (19 mg, 24%). ¹H NMR (CDCl₃, 500 MHz) δ 7.93 (s, 1H), 7.36(m, 2H), 7.29 (s, 1H), 7.26 (m, 3H), 7.13 (s, 1H), 4.30 (m, 2H), 3.18(m, 2H), 2.84 (m, 2H), 2.00 (m, 2H), 1.82 (m, 2H).HRMS calculated forC₁₅H₁₈N₃O [M+H]⁺ 256.1444, found 256.1445.

Step 2: tert-butyl1,3-dioxo-2-((4-phenylpiperidine-1-carbonyl)oxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

tert-Butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Example 1, Step 2, 20 mg, 0.0737 mmol, 1 equiv) was dissolved in 0.4 mldry THF, and triethylamine (38 μl, 5 equiv) and a catalytic amount of4-DMAP were added, followed by(1H-imidazol-1-yl)(4-phenylpiperidin-1-yl)methanone (19 mg, 1 equiv) atroom temperature. The solution was heated at 70° C. for 5 h, andconcentrated to a residue that was separated by prep-TLC (50%EtOAc/hexanes) to yield tert-butyl1,3-dioxo-2-((4-phenylpiperidine-1-carbonyl)oxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylateas a white solid (27 mg, 81%). ¹H NMR (CDCl₃, 400 MHz) δ 7.34 (m, 2H),7.25 (m, 1H), 7.22 (m, 2H), 4.57 (m, 1H), 4.34 (m, 2H), 4.22 (m, 1H),4.11 (m, 2H), 3.14 (m, 1H), 3.08 (m, 1H), 3.02 (m, 1H), 2.86 (m, 2H),2.72 (m, 1H), 1.94 (m, 2H), 1.82 (m, 2H), 1.49 (s, 9H). HRMS calculatedfor C₂₃H₃₀N₄O₆Na [M+Na]⁺ 481.2057, found 481.2056.

Example 28: Preparation of1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-phenylpiperidine-1-carboxylate (Compound 28, JJH322)

tert-Butyl1,3-dioxo-2-((4-phenylpiperidine-1-carbonyl)oxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylatewas Boc-deprotected using methanolic HCl as described for Example 1,Step 4, yielding a white solid (100%). ¹H NMR (CDCl₃, 400 MHz) δ 7.32(m, 2H), 7.25 (m, 1H), 7.22 (m, 2H), 4.89 (m, 1H), 4.33 (m, 2H), 4.21(m, 1H), 3.67 (m, 1H), 3.13 (m, 2H), 3.01 (m, 2H), 2.72 (m, 1H), 1.94(m, 2H), 1.79 (m, 2H), 1.64 (m, 2H). HRMS calculated for C₁₈H₂₃N₄O₄[M+H]⁺ 359.1714, found 359.1715.

Example 29: Preparation of1-(7-(tert-Butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)4-methyl piperidine-1,4-dicarboxylate (Compound 29, JJH238)

Step 1: Methyl 1-(1H-imidazole-1-carbonyl)piperidine-4-carboxylate

Methyl piperidine-4-carboxylatehydrochloride (358 mg, 2 mmol, 1 equiv)was suspended in CH₂Cl₂ (8 ml), triethylamine was added, and thesolution was cooled to 0° C. CDI (405 mg, 1.25 equiv) was added all atonce, and the solution was stirred to room temperature overnight. Theresulting mixture was concentrated under reduced pressure, and separatedby flash chromatography over SiO₂ in 50% acetone/hexanes to yield methyl1-(1H-imidazole-1-carbonyl)piperidine-4-carboxylate as an off-whitesolid (255 mg, 54%). ¹H NMR (CDCl₃, 400 MHz) δ 7.89 (s, 1H), 7.20 (s,1H), 7.11 (s, 1H), 4.04 (m, 2H), 3.99 (s, 3H), 3.24 (m, 2H), 2.66 (m,1H), 2.04 (m, 2H), 1.84 (m, 2H). HRMS calculated for C₁₁H₁₆N₃O₃ [M+H]⁺238.1186, found 238.1185.

Step 2:1-(7-(tert-Butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)4-methyl piperidine-1,4-dicarboxylate

tert-Butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Example 1, Step 2, 7 mg, 0.026 mmol, 1.1 equiv) was dissolved in 0.24ml dry THF, and triethylamine (12 μl, 5 equiv) and a catalytic amount of4-DMAP were added, followed by methyl1-(1H-imidazole-1-carbonyl)piperidine-4-carboxylate (5.4 mg, 1 equiv) atroom temperature. The solution was heated at 70° C. overnight, andconcentrated to a residue that was separated by prep-TLC (75%EtOAc/hexanes) to yield1-(7-(tert-butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)4-methyl piperidine-1,4-dicarboxylate as a white solid (8.3 mg, 73%). ¹HNMR (CDCl₃, 400 MHz) δ 4.56 (m, 1H), 4.17 (m, 1H), 4.07 (m, 3H), 3.98(m, 1H), 3.71 (s, 3H), 3.25 (m, 1H), 3.08 (m, 2H), 2.84 (m, 2H), 2.54(m, 1H), 2.01 (m, 2H), 1.82 (m, 2H), 1.49 (s, 9H). HRMS calculated forC₁₉H₂₈N₄O₈Na [M+Na]⁺ 463.1799, found 463.1799.

Example 30: Preparation of1-(1,3-Dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl) 4-methylpiperidine-1,4-dicarboxylate (Compound 30, JJH257)

1-(7-(tert-Butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)4-methyl piperidine-1,4-dicarboxylate was Boc deprotected usingmethanolic HCl as described for Example 1, Step 4, yielding a whitesolid (100%). ¹H NMR (CDCl₃, 400 MHz) δ 4.94 (m, 1H), 4.28 (m, 1H), 4.06(m, 2H), 3.93 (m, 2H), 3.71 (s, 3H), 3.22 (m, 2H), 3.10 (m, 2H), 2.56(m, 2H), 1.99 (m, 2H), 1.81 (m, 2H). HRMS calculated for C₁₄H₂₁N₄O₆[M+H]⁺ 341.1456, found 341.1455.

Example 31: Preparation of1-(1,3-dioxo-7-(4-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)4-methyl piperidine-1,4-dicarboxylate (Compound 31, ABCS)

The title compound was prepared as described in Example 23, using1-(1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl) 4-methylpiperidine-1,4-dicarboxylate and 3-phenoxybenzaldehyde. (6.8 mg, 87%) ¹HNMR (CDCl₃, 500 MHz) δ 7.36 (m, 2H), 7.28 (m, 1H), 7.11 (m, 1H), 7.01(m, 4H), 6.92 (m, 1H), 4.12 (m, 2H), 4.02 (m, 1H), 3.96 (m, 1H), 3.70(s, 3H), 3.61 (d, J=15 Hz, 1H), 3.53 (d, J=15 Hz, 1H), 3.25 (m, 2H),3.12 (m, 2H), 2.84 (m, 1H), 2.52 (m, 1H), 2.15 (m, 2H), 1.99 (m, 2H),1.81 (m, 2H). HRMS calculated for C₂₇H₃₁N₄O₇ [M+H]⁺ 523.2187, found523.2187.

Example 32: Preparation of7-(tert-butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-ylmorpholine-4-carboxylate (Compound 32, JJH253)

tert-Butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Example 1, Step 2, 29.4 mg, 0.108 mmol, 1 equiv) was dissolved in dryCH₂Cl₂ (0.5 mL), and diisopropylethylamine (45 μl, 3 equiv) and acatalytic amount of 4-DMAP were added, followed by morpholine-4-carbonylchloride (30 μl, ˜2 equiv). The resulting solution was sealed and heatedat 50° C. for 1 h, concentrated to a residue, and separated by prep-TLC(80% EtOAc/hexanes) to yield7-(tert-butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-ylmorpholine-4-carboxylateas a white solid (33 mg, 79%). ¹H NMR (CDC13, 500 MHz) δ 4.54 (m, 1H),4.17 (m, 1H), 4.07 (m, 2H), 3.74 (m, 4H), 3.65 (m, 2H), 3.51 (m, 2H),3.04 (m, 1H), 2.83 (m, 2H), 1.48 (s, 9H). HRMS calculated for C₁₆H₂₅N₄O₇[M+H]⁺ 385.1718, found 385.1710.

Example 33: Preparation of1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-ylmorpholine-4-carboxylate (Compound 33, JJH256)

7-(tert-Butoxycarbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-ylmorpholine-4-carboxylate was Boc deprotected using methanolic HCl asdescribed in Example 1, Step 4, yielding a white solid (100%). ¹H NMR(D₂O, 500 MHz) δ 4.85 (m, 1H), 4.36 (m, 1H), 3.98 (m, 1H), 3.85 (m, 4H),3.73 (m, 2H), 3.65 (m, 1H), 3.58 (m, 2H), 3.52 (m, 1H), 3.36 (m, 1H),3.25 (m, 1H). HRMS calculated for C₁₁H₁₇N₄O₅ [M+H]⁺ 285.1193, found285.1193.

Example 34: Preparation of1,3-dioxo-7-(4-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-ylmorpholine-4-carboxylate (Compound 34, ABC51)

The title compound was prepared as described in Example 23, using1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-ylmorpholine-4-carboxylate and 4-phenoxybenzaldehyde. (18 mg, 64%). ¹H NMR(CDCl₃, 400 MHz) δ 7.37 (m, 2H), 7.27 (m, 2H), 7.13 (m, 1H), 6.98 (m,2H), 6.96 (m, 2H), 4.16 (m, 1H), 4.06 (m, 1H), 3.75 (m, 4H), 3.66 (m,2H), 3.59 (m, 1H), 3.51 (m, 2H), 3.50 (m, 1H), 3.30 (m, 1H), 3.15 (m,2H), 2.89 (m, 2H), 2.10 (m, 1H). HRMS calculated for C₂₄H₂₇N₄O₆ [M+H]⁺467.1925, found 467.1927.

Example 35: Preparation of1,3-dioxo-7-(3-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate (Compound 35, ABC16)

The title compound was prepared as described in Example 23, using1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate and 3-phenoxybenzaldehyde.(22 mg, 69%). ¹H NMR (CDCl₃, 500 MHz) δ 7.36 (m, 2H), 7.28 (m, 3H), 7.11(m, 3H), 7.00 (m, 3H), 6.92 (m, 1H), 4.21 (m, 1H), 4.12 (m, 2H), 4.02(m, 1H), 3.61 (d, J=15 Hz, 1H), 3.53 (d, J=15 Hz, 1H), 3.26 (m, 1H),3.11 (m, 1H), 2.99 (m, 2H), 2.86 (m, 2H), 2.60 (t, J=10 Hz, 2H), 2.14(m, 1H), 1.76 (m, 2H), 1.58 (m, 2H), 1.48 (m, 1H), 1.34 (m, 2H). HRMScalculated for C₃₃H₃₆ClN₄O₅ [M+H]⁺ 603.2369, found 603.2370.

Example 36: Preparation of1,3-dioxo-7-(3-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 36, ABC23)

The title compound was prepared as described in Example 23, using3-phenoxybenzaldehyde in place of formaldehyde. (4.6 mg, 17%). ¹H NMR(CDCl₃, 500 MHz) δ 7.36 (m, 2H), 7.33 (m, 1H), 7.13 (m, 1H), 7.02 (m,4H), 6.92 (m, 3H), 6.86 (d, J=10 Hz, 2H), 4.14 (m, 1H), 4.05 (m, 1H),3.79 (m, 2H), 3.77 (s, 3H), 3.68 (m, 2H), 3.62 (d, J=15 Hz, 1H), 3.54(d, J=15 Hz, 1H), 3.28 (m, 1H), 3.15 (m, 1H), 3.10 (m, 4H), 3.06 (m,1H), 2.87 (m, 1H). HRMS calculated for C₃₁H₃₄N₅O₆ [M+H]⁺ 572.2503, found572.2504.

Example 37: Preparation of1,3-dioxo-7-(4-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 37, ABC34)

The title compound was prepared as described in Example 23, using4-phenoxybenzaldehyde in place of formaldehyde. (6.1 mg, 45%). ¹H NMR(CDCl₃, 500 MHz) δ 7.36 (m, 2H), 7.24 (d, J=10 Hz, 2H), 7.12 (m, 1H),7.04 (d, J=10 Hz, 2H), 6.97 (d, J=10 Hz, 2H), 6.90 (d, J=10 Hz, 2H),6.86 (d, J=8 Hz, 2H), 4.16 (m, 1H), 4.06 (m, 1H), 3.80 (m, 2H), 3.78 (s,3H), 3.68 (m, 2H), 3.62 (d, J=15 Hz, 1H), 3.53 (d, J=15 Hz, 1H), 3.30(m, 1H), 3.18 (m, 1H), 3.11 (m, 4H), 3.04 (m, 1H), 2.89 (m, 1H). HRMScalculated for C₃₁H₃₄N₅O₆ [M+H]⁺ 572.2503, found 572.2504.

Example 38: Preparation of1,3-dioxo-7-(4-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-phenylpiperidine-1-carboxylate (Compound 38, ABC47)

The title compound was prepared as described in Example 23, using1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-phenylpiperidine-1-carboxylate and 4-phenoxybenzaldehyde. (19 mg,62%). ¹H NMR (CDCl₃, 500 MHz) δ 7.36 (m, 4H), 7.28 (m, 5H), 7.11 (m,1H), 7.04 (m, 2H), 6.97 (m, 2H), 4.36 (m, 1H), 4.28 (m, 1H), 4.17 (m,1H), 4.08 (m, 1H), 3.63 (d, J=15 Hz, 1H), 3.54 (d, J=15 Hz, 1H), 3.31(m, 1H), 3.16 (m, 2H), 3.01 (m, 1H), 2.90 (m, 1H), 2.74 (m, 1H), 2.18(m, 2H), 1.91 (m, 2H), 1.80 (m, 2H). HRMS calculated for C₃₁H₃₃N₄O₅[M+H]⁺ 541.2445, found 541.2448.

Example 39: Preparation of1,3-dioxo-7-(4-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carboxylate (Compound 39,ABC45)

Step 1: tert-Butyl4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carboxylate

tert-Butyl 4-(4-hydroxyphenyl)piperazine-1-carboxylate (100 mg, 360μmol, 1 equiv) was suspended in 0.75 mL DMF, and propargyl bromide (80%by weight in toluene, 67 μL, 440 μmol, 1.25 equiv) and potassiumcarbonate (75 mg, 540 μmol, 1.5 equiv) were added. The resulting mixturewas stirred at 60° C. for 18 h, cooled to room temperature, concentratedunder a stream of nitrogen, and separated by prep-TLC (25%EtOAc/hexanes) to yield an off-white solid (62 mg, 54%). ¹H NMR (CDCl3,500 MHz) δ 6.91 (m, 4H), 4.64 (d, J=2.4 Hz, 2H), 3.57 (t, J=5.0 Hz, 4H),3.02 (t, J=5.0 Hz, 4H), 2.50 (t, J=2.4 Hz, 1H), 1.48 (s, 9H). HRMScalculated for C₁₈H₂₅N₂O₃ [M+H]⁺ 317.186, found 317.1858.

Step 2: 1-(4-(Prop-2-yn-1-yloxy)phenyl)piperazine hydrochloride

tert-Butyl 4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carboxylate wasBoc deprotected using methanolic HCl as described for Example 1, Step 4,yielding an off-white solid (100%). ¹H NMR (D₂O, 500 MHz) δ 7.43 (m,2H), 7.18 (m, 2H), 4.83 (d, J=2.3 Hz, 2H), 3.76 (m, 4H), 3.67 (m, 4H),2.98 (t, J=2.4 Hz, 1H). HRMS calculated for C₁₃H₁₇N₂O [M+H]⁺ 217.1335,found 217.1335.

Step 3:(1H-Imidazol-1-yl)(4-(4-(prop-2-yn-1-yloxy)phenyl)piperazin-1-yl)methanone

1-(4-(Prop-2-yn-1-yloxy)phenyl)piperazine hydrochloride (49 mg, 196μmol, 1 equiv) was taken up in CH₂Cl₂ (2 mL), and triethylamine (27 μL,196 μmol, 1 equiv) and CDI (48 mg, 294 μmol, 1.5 equiv) were added. Theresulting mixture was stirred at room temperature overnight,concentrated and separated by flash chromatography over SiO₂ in 60%EtOAc/hexanes to yield the corresponding imidazole urea (47 mg, 77%). ¹HNMR (CDCl₃, 500 MHz) δ 7.92 (s, 1H), 7.23 (t, J=1.4 Hz, 1H), 7.13 (br m,1H), 6.93 (m, 4H), 4.65 (d, J=2.4 Hz, 2H), 3.77 (m, 4H), 3.15 (m, 4H),2.51 (t, J=2.3 Hz, 1H). HRMS calculated for C₁₇H₁₉N₄O₂ [M+H]⁺ 311.1502,found 311.1504.

Step 4: tert-Butyl1,3-dioxo-2-((4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carbonyl)oxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

tert-Butyl2-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(Example 1, Step 2, 9.5 mg, 35.2 μmol, 1.1 equiv) was dissolved in 0.35mL THF, and triethylamine (13 μl, 96 μmol, 3 equiv) was added, followedby(1H-imidazol-1-yl)(4-(4-(prop-2-yn-1-yloxy)phenyl)piperazin-1-yl)methanone(10 mg, 32 μmol, 1 equiv) at room temperature. The solution was stirredand heated at 70° C. for 2 h, concentrated to a residue, and separatedby prep-TLC (25% EtOAc/hexanes) to yield an off-white solid (14 mg,85%). ¹H NMR (CDCl₃, 500 MHz) δ 6.92 (m, 4H), 4.65 (d, J=2.3 Hz, 2H),4.56 (s, 1H), 4.19 (s, 1H), 4.08 (m, 2H), 3.80 (s, 2H), 3.67 (s, 2H),3.13 (s, 4H), 3.05 (t, J=12.0 Hz, 1H), 2.85 (s, 2H), 2.51 (t, J=2.4 Hz,1H), 1.49 (s, 9H). HRMS calculated for C₂₅H₃₂N₅O₇ [M+H]⁺ 514.2296, found514.2296.

Step 5:1,3-Dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carboxylatehydrochloride

tert-Butyl1,3-dioxo-2-((4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carbonyl)oxy)hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylatewas Boc deprotected using methanolic HCl as described for Example 1,Step 4, yielding an off-white solid (100%). ¹H NMR (D₂O, 500 MHz) δ 7.58(m, 2H), 7.25 (m, 2H), 4.87 (m, 3H), 4.36 (m, 1H), 4.16 (s, 2H), 3.99(m, 3H), 3.80 (br s, 4H), 3.64 (dd, J=13.3, 3.5 Hz, 1H), 3.54 (ddd,J=15.0, 12.8, 3.7 Hz, 1H), 3.39 (t, J=12.5 Hz, 1H), 3.28 (td, J=12.9,4.5 Hz, 1H), 3.00 (t, J=2.4 Hz, 1H). HRMS calculated for C₂₀H₂₄N₅O₅[M+H]⁺ 414.1772, found 414.1771.

Step 6:1,3-Dioxo-7-(4-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carboxylate

1,3-Dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carboxylate hydrochloride(12 mg, 27 μmol, 1 equiv) was suspended in CH₂Cl₂ (270 μL), and4-phenoxy benzaldehyde (22 mg, 109 μmol, 4 equiv) was added followed byNaBH(OAc)₃ (12.2 mg, 54 μmol, 2 equiv). The resulting mixture wasstirred at room temperature overnight, concentrated and separated byprep-TLC (50% EtOAc/hexanes) to yield1,3-dioxo-7-(4-phenoxybenzyl)hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-yn-1-yloxy)phenyl)piperazine-1-carboxylate as a clear oil(11.6 mg, 72%). ¹H NMR (CDCl₃, 500 MHz) δ 7.35 (m, 2H), 7.24 (m, 3H),7.12 (t, J=7.3 Hz, 1H), 7.03 (m, 2H), 6.96 (m, 2H), 6.92 (m, 3H), 4.65(br s, J=2.7, 1.5 Hz, 2H), 4.15 (d, J=10.5 Hz, 1H), 4.05 (d, J=13.3 Hz,1H), 3.79 (s, 2H), 3.67 (s, 2H), 3.60 (d, J=13.0 Hz, 1H), 3.51 (d,J=12.7 Hz, 1H), 3.29 (d, J=9.5 Hz, 1H), 3.13 (s, 5H), 2.88 (d, J=11.1Hz, 1H), 2.51 (s, 1H), 2.14 (br s, 2H). HRMS calculated for C₃₃H₃₄N₅O₆[M+H]⁺ 596.2503, found 596.2507.

Example 40: Preparation of(S)-7-(4-bromobenzyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate(Compound 40)

7-(4-Bromobenzyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate(Example 13, 95.7 mg, 0.162 mmol, 1.00 equiv) was separated by SFC usingthe following gradient conditions: Column: Phenomenex Lux 5u Cellulose-35*25 cm, 5 um; Mobile Phase A: CO₂ (60%), Mobile Phase B: acetonitrile(40%); Flow rate: 140 mL/min; Detector, UV 220 nm; RT1: 5.2 min; RT2:6.55 min. SFC separation resulted in 75.3 mg (79% yield) of(S)-7-(4-bromobenzyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylateas an off-white semi-solid. ¹H NMR (CDCl₃, 300 MHz) δ 7.46 (d, J=8.4 Hz,2H), 7.19-7.24 (m, 4H), 7.09 (d, J=8.4 Hz, 2H), 4.02-4.22 (m, 4H), 3.56(br, 2H), 3.16-3.34 (m, 2H), 2.85-3.01 (m, 3H), 2.60 (t, J=7.8 Hz, 2H),2.17 (br, 2H), 1.75-1.79 (m, 2H), 1.54-1.61 (m, 2H), 1.25-1.33 (m, 3H).LCMS (ESI, m/z): 591 [M+H]⁺.

Or alternatively:

Step 1: (S)-tert-Butyl2-(benzyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

A 250-mL round-bottom flask was charged with CDI (16.2 g, 99.9 mmol,1.00 equiv), CH₂Cl₂ (120 mL), O-benzylhydroxylamine (12.3 g, 99.9 mmol,1.00 equiv). NMM (40.4 g, 399 mmol, 4.00 equiv) was added. The mixturewas stirred at room temperature for 2 h.(S)-4-(tert-Butoxycarbonyl)piperazine-2-carboxylic acid (23.0 g, 99.9mmol, 1.00 equiv) was added. The resulting solution was stirredovernight at room temperature and quenched with water (100 mL). Theresulting solution was extracted with CH₂Cl₂ (3×200 mL) and the organiclayers were combined, washed with water (3×50 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure. The residuewas chromatographed on a silica gel column with EtOAc/petroleum ether(1/3) to provide 13.7 g (38% yield) of (S)-tert-butyl2-(benzyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylateas a white solid. LCMS (ESI, m/z): 362 [M+H]⁺.

Step 2: (S)-tert-Butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

A 25-mL round-bottom flask was charged with (S)-tert-butyl2-(benzyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(1.00 g, 2.77 mmol, 1.00 equiv), palladium carbon (200 mg), ethanol (8mL), EtOAc (2 mL). H₂ (g) was introduced in. The resulting solution wasstirred overnight at room temperature. The solids were filtered out. Theresulting mixture was concentrated to provide 700 mg (93% yield) of(S)-tert-butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate asa white solid. LCMS (ESI, m/z): 272 [M+H]⁺.

Step 3: 4-[2-(4-Chlorophenyl)ethyl]piperidine-1-carbonyl chloride

A 25-mL round-bottom flask was charged with triphosgene (110 mg, 0.370mmol, 0.50 equiv), CH₂Cl₂ (5 mL). The mixture was cooled to 0° C.4-[2-(4-Chlorophenyl)ethyl]piperidine (165 mg, 0.737 mmol, 1.00 equiv)was added. N,N-Diisopropylethylamine (381 mg, 2.95 mmol, 4.00 equiv) wasadded dropwise. The resulting solution was stirred for 2 h at roomtemperature and quenched with water (10 mL). The resulting solution wasextracted with CH₂Cl₂ (3×30 mL) and the organic layers were combined,washed with water (3×10 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated to provide 210 mg (99% yield) of4-[2-(4-chlorophenyl)ethyl]piperidine-1-carbonyl chloride as yellow oil.LCMS (ESI, m/z): 286 [M+H]⁺.

Step 4: (S)-tert-Butyl2-(4-(4-chlorophenethyl)piperidine-1-carbonyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

A 25-mL round-bottom flask was charged with4-[2-(4-chlorophenyl)ethyl]piperidine-1-carbonyl chloride (210 mg, 0.734mmol, 1.00 equiv), (S)-tert-butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(240 mg, 0.886 mmol, 1.20 equiv), 4-DMAP (18.0 mg, 0.148 mmol, 0.20equiv), NMM (224 mg, 2.22 mmol, 3.00 equiv), CH₂Cl₂ (5 mL). Theresulting solution was stirred overnight at room temperature andquenched with water (10 mL). The resulting solution was extracted withCH₂Cl₂ (3×30 mL) and the organic layers were combined, washed with water(3×10 mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure. The residue was chromatographed on a silica gel columnwith EtOAc/petroleum ether (1/1) to provide 250 mg (65% yield) of(S)-tert-butyl2-(4-(4-chlorophenethyl)piperidine-1-carbonyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylateas a colorless solid. LCMS (ESI, m/z): 521 [M+H]⁺.

Step 5: (S)-1,3-Dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate

A 25-mL round-bottom flask was charged with (S)-tert-butyl2-(4-(4-chlorophenethyl)piperidine-1-carbonyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(250 mg, 0.480 mmol, 1.00 equiv), CH₂Cl₂ (5 mL). The mixture was cooledto 0° C. Trifluoroacetic acid (1 mL) was added. The resulting solutionwas stirred for 3 h at room temperature and concentrated to provide 200mg (99% yield) of(S)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylateas a yellow solid. LCMS (ESI, m/z): 421 [M+H]⁺.

Step 6:(S)-7-(4-Bromobenzyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate

A 25-mL round-bottom flask was charged with(S)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylate(200 mg, 0.475 mmol, 1.00 equiv), 4-bromobenzaldehyde (105 mg, 0.568mmol, 1.20 equiv), 1,2-dichloroethane (5 mL), triethylamine (144 mg,1.42 mmol, 3.00 equiv). The mixture was stirred at room temperature for30 min. NaBH(OAc)₃ (302 mg, 1.42 mmol, 3.00 equiv) was added. Theresulting solution was stirred for 3 h at room temperature and quenchedwith water (10 mL). The resulting solution was extracted with CH₂Cl₂(3×30 mL) and the organic layers were combined, washed with water (3×10mL), dried over anhydrous Na₂SO₄, filtered and concentrated. The residuewas chromatographed on a silica gel column with MeOH/CH₂Cl₂ (2/98). Thecrude product (250 mg) was purified by preparative HPLC using thefollowing gradient conditions: 20% CH₃CN/80% Phase A increasing to 80%CH₃CN over 10 min, then to 100% CH₃CN over 0.1 min, holding at 100%CH₃CN for 1.9 min, then reducing to 20% CH₃CN over 0.1 min, and holdingat 20% for 1.9 min, on a Waters 2767-5 Chromatograph. Column: XbridgePrep C18, 19*150 mm 5 um; Mobile phase: Phase A: water; Phase B: CH₃CN;Detector, UV220 & 254 nm. Purification resulted in 121.5 mg (43% yield)of(S)-7-(4-bromobenzyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperidine-1-carboxylateas a white solid. ¹H NMR (CDCl₃, 300 MHz) δ 7.46 (d, J=8.4 Hz. 2H),7.18-7.24 (m, 4H), 7.09 (d, J=8.4 Hz, 2H), 4.02-4.22 (m, 4H), 3.49-3.60(m, 2H), 3.14-3.25 (m, 2H), 2.82-3.01 (m, 3H), 2.60 (t, J=7.8 Hz, 2H),2.13-2.16 (m, 2H), 1.75-1.79 (m, 2H), 1.54-1.61 (m, 3H), 1.25-1.38 (m,2H). LCMS (ESI, m/z): 591 [M+H]⁺.

Example 41:(S)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperazine-1-carboxylate (Compound 41)

Step 1.(S)-2-(benzyloxy)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione

A 250-mL round-bottom flask was charged with (S)-tert-butyl2-(benzyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(7.22 g, 20.0 mmol, 1.00 equiv), MeOH (70 mL), HCl (14 mL). Theresulting solution was stirred overnight at room temperature andconcentrated to provide 5.00 g (96% yield) of(S)-2-(benzyloxy)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione as awhite solid. LCMS (ESI, m/z): 262 [M+H]⁺.

Step 2.(S)-2-(benzyloxy)-7-(morpholine-4-carbonyl)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione

A 250-mL round-bottom flask was charged with(S)-2-(benzyloxy)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione (5.00g, 19.1 mmol, 1.00 equiv), morpholine-4-carbonyl chloride (3.45 g, 23.1mmol, 1.20 equiv), 4-DMAP (467 mg, 3.82 mmol, 0.20 equiv), triethylamine(5.80 g, 57.3 mmol, 3.00 equiv), and THF (50 mL). The resulting solutionwas stirred for 2 h at room temperature and diluted with water (50 mL).The resulting solution was extracted with CH₂Cl₂ (3×100 mL) and theorganic layers were combined, washed with water (3×50 mL), dried overanhydrous Na₂SO₄, filtered and concentrated. The residue waschromatographed on a silica gel column with EtOAc/petroleum ether (99/1)to provide 6.80 g (95% yield) of(S)-2-(benzyloxy)-7-(morpholine-4-carbonyl)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dioneas a white solid. LCMS (ESI, m/z): 375 [M+H]⁺.

Step 3:(S)-2-hydroxy-7-(morpholine-4-carbonyl)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione

A 250-mL round-bottom flask was charged with(S)-2-(benzyloxy)-7-(morpholine-4-carbonyl)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione(7.20 g, 19.2 mmol, 1.00 equiv), palladium carbon (1.50 g), ethanol (56mL), EtOAc (14 mL). H₂ (g) was introduced into the reaction mixture. Theresulting solution was stirred overnight at room temperature. The solidswere filtered out. The resulting mixture was concentrated to provide5.30 g (97% yield) of(S)-2-hydroxy-7-(morpholine-4-carbonyl)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dioneas a white semi-solid. LCMS (ESI, m/z): 285 [M+H]⁺.

Step 5.(S)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperazine-1-carboxylate

A 25-mL round-bottom flask was charged with4-[2-(4-chlorophenyl)ethyl]piperazine-1-carbonyl chloride (117 mg, 0.408mmol, 1.00 equiv),(S)-2-hydroxy-7-(morpholine-4-carbonyl)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione (116 mg, 0.407 mmol, 1.00 equiv), 4-DMAP (9.95 mg, 0.0816mmol, 0.20 equiv), N,N-diisopropylethylamine (158 mg, 1.22 mmol, 3.00equiv), CH₂Cl₂ (5 mL). The resulting solution was stirred overnight atroom temperature and concentrated. The crude product (200 mg) waspurified by preparative HPLC using the following gradient conditions:20% CH₃CN/80% Phase A increasing to 80% CH₃CN over 10 min, then to 100%CH₃CN over 0.1 min, holding at 100% CH₃CN for 1.9 min, then reducing to20% CH₃CN over 0.1 min, and holding at 20% for 1.9 min, on a Waters2767-5 Chromatograph. Column: Xbridge Prep C18, 19*150 mm 5 um; Mobilephase: Phase A: water; Phase B: CH₃CN; Detector, UV220 & 254 nm. Theresidue was chromatographed on a silica gel column with MeOH/CH₂Cl₂(4/96) to provide 31.6 mg (14% yield) of(S)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chlorophenethyl)piperazine-1-carboxylate as a white solid. ¹H NMR(CDCl₃, 300 MHz) δ 7.25-7.27 (m, 2H), 7.12-7.15 (m, 2H), 4.22-4.27 (m,1H), 4.01-4.10 (m, 2H), 3.65-3.70 (m, 7H), 3.59 (br, 2H), 3.32-3.34 (m,4H), 3.10-3.20 (m, 1H), 2.96-3.05 (m, 2H), 2.80 (br, 2H), 2.63 (br, 6H).LCMS (ESI, m/z): 535 [M+H]⁺.

Example 42: Preparation of(S)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 42)

7-((1-Benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Example 26, 320 mg, 0.554mmol, 1.00 equiv) was separated by SFC using the following gradientconditions: Column: CHIRALPAK AD-H SFC 5*25 cm, 5 μm; Mobile Phase A:CO₂: (50%), Mobile Phase B: isopropanol (0.1% diethylamine) (50%); Flowrate: 190 mL/min; Detector, UV 220 nm; RT1: 6.41 min; RT2: 8.50 min. SFCseparation resulted in 136.5 mg (42% yield) of(S)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylateas a brown semi-solid. ¹H NMR (CDCl₃, 300 MHz) δ 7.28-7.34 (m, 5H),6.83-6.92 (m, 4H), 4.10-4.15 (m, 1H), 3.99-4.04 (m, 1H), 3.78 (m, 5H),3.68 (m, 2H), 3.55 (br, 2H), 3.22-3.27 (m, 1H), 3.06-3.16 (m, 5H), 2.92(br, 2H), 2.77-2.81 (m, 1H), 2.27 (d, J=6.9 Hz, 2H), 2.00-2.14 (m. 4H),1.67-1.76 (m, 2H), 1.51 (br, 1H), 1.26-1.29 (m, 2H). LCMS (ESI, m/z):577 [M+H]⁺.

Or alternatively:

Step 1: 4-(4-Methoxyphenyl)piperazine-1-carbonyl chloride

A 25-mL round-bottom flask was charged with triphosgene (387 mg, 1.30mmol, 0.50 equiv), CH₂Cl₂ (5 mL). The mixture was cooled to 0° C.1-(4-Methoxyphenyl)piperazine (500 mg, 2.60 mmol, 1.00 equiv) was added.N,N-Diisopropylethylamine (1.34 g, 10.4 mmol, 4.00 equiv) was added. Theresulting solution was stirred for 2 h at room temperature and quenchedwith water (10 mL). The resulting solution was extracted with CH₂Cl₂(3×30 mL) and the organic layers were combined, washed with water (3×10mL), dried over anhydrous Na₂SO₄, filtered and concentrated to provide600 mg (91% yield) of 4-(4-methoxyphenyl)piperazine-1-carbonyl chlorideas yellow oil. LCMS (ESI, m/z): 255 [M+H]⁺.

Step 2: (S)-tert-Butyl2-(4-(4-methoxyphenyl)piperazine-1-carbonyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

A 25-mL round-bottom flask was charged with (S)-tert-butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(638 mg, 2.35 mmol, 1.00 equiv),4-(4-methoxyphenyl)piperazine-1-carbonyl chloride (600 mg, 2.36 mmol,1.00 equiv), NMM (713 mg, 7.05 mmol, 3.00 equiv), 4-DMAP (57.4 mg, 0.470mmol, 0.20 equiv), CH₂Cl₂ (5 mL). The resulting solution was stirredovernight at room temperature and quenched with water (10 mL). Theresulting solution was extracted with CH₂Cl₂ (3×30 mL) and the organiclayers were combined, washed with water (3×10 mL), dried over anhydrousNa₂SO₄, filtered and concentrated. The residue was chromatographed on asilica gel column with EtOAc/petroleum ether (4/6) to provide 590 mg(51% yield) of (S)-tert-butyl2-(4-(4-methoxyphenyl)piperazine-1-carbonyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylateas an off-white solid. LCMS (ESI, m/z): 490 [M+H]⁺.

Step 3:(S)-1,3-Dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate

A 25-mL round-bottom flask was charged with (S)-tert-butyl2-(4-(4-methoxyphenyl)piperazine-1-carbonyloxy)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(490 mg, 1.00 mmol, 1.00 equiv), CH₂Cl₂ (8 mL), trifluoroacetic acid (2mL). The resulting solution was stirred overnight at room temperatureand concentrated to provide 460 mg (crude) of(S)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate as a brown solid. LCMS (ESI,m/z): 390 [M+H]⁺.

Step 4:(S)-7-((1-Benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate

A 25-mL round-bottom flask was charged with1-benzylpiperidine-4-carbaldehyde (192 mg, 0.946 mmol, 3.00 equiv),(S)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (123 mg, 0.316 mmol, 1.00equiv), 1,2-dichloroethane (5 mL). Triethylamine (48.0 mg, 0.480 mmol,1.50 equiv) was added. The mixture was stirred at room temperature for30 min. NaBH(OAc)₃ (201 mg, 0.948 mmol, 3.00 equiv) was added. Theresulting solution was stirred overnight at room temperature andquenched with water (10 mL). The resulting solution was extracted withCH₂Cl₂ (3×30 mL) and the organic layers were combined, washed with water(3×10 mL), dried over anhydrous Na₂SO₄, filtered and concentrated. Theresidue was chromatographed on a silica gel column with EtOAc/petroleumether (95/5). The crude product (300 mg) was purified by preparativeHPLC using the following gradient conditions: 20% CH₃CN/80% Phase Aincreasing to 80% CH₃CN over 10 min, then to 100% CH₃CN over 0.1 min,holding at 100% CH₃CN for 1.9 min, then reducing to 20% CH₃CN over 0.1min, and holding at 20% for 1.9 min, on a Waters 2767-5 Chromatograph.Column: Xbridge Prep C18, 19*150 mm 5 um; Mobile phase: Phase A: water;Phase B: CH₃CN; Detector, UV220 & 254 nm. Purification resulted in 128.7mg (71% yield) of(S)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylateas an off-white semi-solid. ¹H NMR (CDCl₃, 300 MHz) δ 7.27-7.35 (m, 5H),6.83-6.92 (m, 4H), 4.10-4.15 (m, 1H), 3.99-4.04 (m, 1H), 3.85 (br, 5H),3.60-3.78 (m, 4H), 3.22-3.27 (m, 1H), 3.06-3.16 (m, 5H), 2.98 (br, 2H),2.77-2.80 (m, 1H), 2.28 (d, J=6.6 Hz, 2H), 2.07-2.14 (m. 4H), 1.68-1.78(m, 3H), 1.36 (br, 2H). LCMS (ESI, m/z): 577 [M+H]⁺.

Example 43: Preparation of(R)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Compound 43)

7-((1-Benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate (Example 26, 320 mg, 0.554mmol, 1.00 equiv) was separated by SFC using the following gradientconditions: Column: CHIRALPAK AD-H SFC 5*25 cm, 5 μm; Mobile Phase A:CO₂: (50%), Mobile Phase B: isopropanol (0.1% diethylamine) (50%); Flowrate: 190 mL/min; Detector, UV 220 nm; RT1: 6.41 min; RT2: 8.50 min. SFCseparation resulted in 86.1 mg (26% yield) of(R)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-methoxyphenyl)piperazine-1-carboxylate as a brown semi-solid. ¹HNMR (CDCl₃, 300 MHz) δ 7.29-7.32 (m, 5H), 6.83-6.92 (m, 4H), 4.10-4.15(m, 1H), 3.99-4.04 (m, 1H), 3.78 (br, 5H), 3.68 (br, 2H), 3.54 (br, 2H),3.22-3.27 (m, 1H), 3.06-3.16 (m, 5H), 2.92 (d, J=9.0 Hz, 2H), 2.77-2.81(m, 1H), 2.27 (d, J=6.9 Hz, 2H), 1.99-2.14 (m. 4H), 1.67-1.76 (m, 2H),1.46-1.51 (m, 1H), 1.26-1.29 (m, 2H). LCMS (ESI, m/z): 577 [M+H]⁺.

Alternatively, the title compound was synthesized as described inExample 42 (Step 1-4) using (R)-tert-butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate asthe starting material. ¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.36 (m, 5H),6.83-6.92 (m, 4H), 3.99-4.15 (m, 2H), 3.49-3.85 (m, 9H), 2.77-3.27 (m,9H), 2.28 (d, J=6.3 Hz, 2H), 2.05-2.15 (m, 4H), 1.68-1.79 (m, 3H),1.23-1.33 (m, 2H). LCMS (ESI, m/z): 577 [M+H]⁺.

Example 44: Preparation of(S)-1,3-dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate (Compound 44)

1,3-Dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate(Example 39, 435 mg, 0.730 mmol, 1.00 equiv) was separated by SFC usingthe following gradient conditions: Column: Chiralpak IC 2*25 cm, 5 um;Mobile Phase A: CO₂ (50%), Mobile Phase B: acetonitrile (50%); Flowrate: 40 mL/min; Detector, UV 220 nm; RT1: 8.95 min; RT2: 10.84 min. SFCseparation resulted in 274.4 mg (63% yield) of(S)-1,3-dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate as a white solid.¹H NMR (CDCl₃, 300 MHz) δ 7.25-7.38 (m, 3H), 6.89-7.14 (m, 10H), 4.65(d, J=2.4 Hz, 2H), 4.04-4.17 (m, 2H), 3.49-3.80 (m, 6H), 3.13-3.31 (m,6H), 2.87-2.91 (m, 1H), 2.51 (t, J=2.4 Hz, 1H), 2.13-2.16 (m, 2H). LCMS(ESI, m/z): 596 [M+H]⁺.

Alternatively, the title compound was prepared as follows.

Step 1: tert-Butyl4-[4-(prop-2-yn-1-yloxy)phenyl]piperazine-1-carboxylate

A 25-mL round-bottom flask was charged with 3-bromoprop-1-yne (428 mg,3.60 mmol, 1.00 equiv), tert-butyl4-(4-hydroxyphenyl)piperazine-1-carboxylate (1.00 g, 3.59 mmol, 1.00equiv), cesium carbonate (3.52 g, 10.8 mmol, 3.00 equiv),N,N-dimethylformamide (10 mL). The resulting solution was stirredovernight at 50° C. and diluted with water (10 mL). The resultingsolution was extracted with CH₂Cl₂ (3×30 mL) and the organic layers werecombined, washed with water (3×10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated. The residue was chromatographed on a silicagel column with EtOAc/petroleum ether (15/85) to provide 1.04 g (91%yield) of tert-butyl4-[4-(prop-2-yn-1-yloxy)phenyl]piperazine-1-carboxylate as yellow oil.LCMS (ESI, m/z): 317 [M+H]⁺.

Step 2: 1-[4-(Prop-2-yn-1-yloxy)phenyl]piperazine

A 100-mL round-bottom flask was charged with tert-butyl4-[4-(prop-2-yn-1-yloxy)phenyl]piperazine-1-carboxylate (930 mg, 2.94mmol, 1.00 equiv), CH₂Cl₂ (10 mL). The mixture was cooled to 0° C.Trifluoroacetic acid (2 mL) was added dropwise. The resulting solutionwas stirred for 3 h at room temperature and concentrated to provide 690mg (crude) of 1-[4-(prop-2-yn-1-yloxy)phenyl]piperazine as a brownsolid. LCMS (ESI, m/z): 217 [M+H]⁺.

Step 3: 4-[4-(Prop-2-yn-1-yloxy)phenyl]piperazine-1-carbonyl chloride

A 25-mL round-bottom flask was charged with triphosgene (474 mg, 1.60mmol, 0.50 equiv), CH₂Cl₂ (5 mL). The mixture was cooled to 0° C.1-[4-(Prop-2-yn-1-yloxy)phenyl]piperazine (690 mg, 3.19 mmol, 1.00equiv) was added. N,N-Diisopropylethylamine (1.65 g, 12.8 mmol, 4.00equiv) was added dropwise. The resulting solution was stirred for 2 h atroom temperature and quenched with water (10 mL). The resulting solutionwas extracted with CH₂Cl₂ (3×30 mL) and the organic layers werecombined, washed with water (3×10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated to provide 880 mg (99% yield) of4-[4-(prop-2-yn-1-yloxy)phenyl]piperazine-1-carbonyl chloride as yellowoil. LCMS (ESI, m/z): 279 [M+H]⁺.

Step 4: (S)-tert-Butyl1,3-dioxo-2-(4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carbonyloxy)-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate

A 25-mL round-bottom flask was charged with (S)-tert-butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(855 mg, 3.15 mmol, 1.00 equiv),4-[4-(prop-2-yn-1-yloxy)phenyl]piperazine-1-carbonyl chloride (880 mg,3.16 mmol, 1.00 equiv), NMM (956 mg, 9.45 mmol, 3.00 equiv), 4-DMAP(77.0 mg, 0.631 mmol, 0.20 equiv), CH₂Cl₂ (10 mL). The resultingsolution was stirred overnight at room temperature and quenched withwater (10 mL). The resulting solution was extracted with CH₂Cl₂ (3×30mL) and the organic layers were combined, washed with water (3×10 mL),dried over anhydrous Na₂SO₄, filtered and concentrated. The residue waschromatographed on a silica gel column with EtOAc/petroleum ether (3/1)to provide 1.00 g (62% yield) of (S)-tert-butyl1,3-dioxo-2-(4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carbonyloxy)-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylateas an off-white solid. LCMS (ESI, m/z): 514 [M+H]⁺.

Step 5:(S)-1,3-Dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate

A 25-mL round-bottom flask was charged with (S)-tert-butyl1,3-dioxo-2-(4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carbonyloxy)-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate(900 mg, 1.75 mmol, 1.00 equiv), CH₂Cl₂ (10 mL). The mixture was cooledto 0° C. Trifluoroacetic acid (2 mL) was added dropwise. The resultingsolution was stirred for 3 h at room temperature and concentrated toprovide 720 mg (99% yield) of(S)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylateas a brown solid. LCMS (ESI, m/z): 414 [M+H]⁺.

Step 6:(S)-1,3-Dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate

A 50-mL round-bottom flask was charged with 4-phenoxybenzaldehyde (414mg, 2.09 mmol, 1.20 equiv),(S)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate(720 mg, 1.74 mmol, 1.00 equiv), 1,2-dichloroethane (10 mL),triethylamine (528 mg, 5.22 mmol, 3.00 equiv). The mixture was stirredat room temperature for 30 min. NaBH(OAc)₃ (1.11 g, 5.24 mmol, 3.00equiv) was added. The resulting solution was stirred overnight at roomtemperature and quenched with water (10 mL). The resulting solution wasextracted with CH₂Cl₂ (3×30 mL) and the organic layers were combined,washed with water (3×10 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated. The residue was chromatographed on a silica gel columnwith EtOAc/petroleum ether (9/1). The crude product was triturated inEtOAc/hexane (9/1) to provide 399.8 mg (39% yield) of(S)-1,3-dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylateas a white solid. ¹H NMR (CDCl₃, 300 MHz) δ 7.36 (t, J=7.8 Hz, 2H), 7.26(br, 2H), 7.12 (t, J=7.4 Hz, 1H), 6.89-7.05 (m, 8H), 4.65 (d, J=2.4 Hz,2H), 4.04-4.15 (m, 2H), 3.54-3.89 (m, 6H), 2.89-3.31 (m, 7H), 2.51 (t,J=2.4 Hz, 1H), 2.13-2.16 (m, 2H). LCMS (ESI, m/z): 596 [M+H]⁺.

Example 45: Preparation of(R)-1,3-dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate (Compound 45)

1,3-Dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate(Example 39, 435 mg, 0.730 mmol, 1.00 equiv) was separated by SFC usingthe following gradient conditions: Column: Chiralpak IC 2*25 cm, 5 um;Mobile Phase A: CO₂ (50%), Mobile Phase B: acetonitrile (50%); Flowrate: 40 mL/min; Detector, UV 220 nm; RT1: 8.95 min; RT2: 10.84 min. SFCseparation resulted in 45.2 mg (10% yield) of(R)-1,3-dioxo-7-(4-phenoxybenzyl)-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate as an off-whitesolid. ¹H NMR (CDCl₃, 300 MHz) δ 7.26-7.38 (m, 4H), 6.93-7.15 (m, 9H),4.65 (d, J=2.1 Hz, 2H), 4.04-4.16 (m, 2H), 3.54-3.90 (m, 6H), 3.14-3.29(m, 6H), 2.90-2.92 (m, 1H), 2.51 (t, J=2.4 Hz, 1H), 2.17 (br, 2H). LCMS(ESI, m/z): 596 [M+H]⁺.

Alternatively, the title compound was synthesized as described inExample 44 (Step 1-6) using (R)-tert-butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate asthe starting material. ¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.36 (m, 5H),6.83-6.92 (m, 4H), 3.99-4.15 (m, 2H), 3.49-3.85 (m, 9H), 2.77-3.27 (m,9H), 2.28 (d, J=6.3 Hz, 2H), 2.05-2.15 (m, 4H), 1.68-1.79 (m, 3H),1.23-1.33 (m, 2H). LCMS (ESI, m/z): 577 [M+H]⁺.

Example 46: Preparation of(S)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate (Compound 46)

7-((1-Benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate (300 mg, 0.50 mmol,1.00 equiv) was separated by chiral-HPLC using the following gradientconditions: Column: Chiralpak IA 2*25 cm, 5 um; Mobile Phase A:acetonitrile (95%), Mobile Phase B: CH₂Cl₂ (5%); Flow rate: 20 mL/min;Detector, UV 254 nm; RT1: 11.50 min; RT2: 19.0 min. Chiral-HPLCseparation resulted in 128.5 mg (43% yield) of(S)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate as a white solid.¹H NMR (CDCl₃, 300 MHz) δ 7.31-7.33 (m, 5H), 6.88-6.95 (m, 4H), 4.65 (d,J=2.4 Hz, 2H), 4.10-4.15 (m, 1H), 3.99-4.04 (m, 1H), 3.80 (br, 2H),3.58-3.67 (m, 4H), 3.22-3.27 (m, 1H), 2.95-3.14 (m, 7H), 2.77-2.81 (m,1H), 2.51 (t, J=2.4 Hz, 1H), 2.28 (d, J=6.9 Hz, 2H), 2.03-2.14 (m, 4H),1.68-1.78 (m, 3H), 1.32-1.34 (m, 2H). LCMS (ESI, m/z): 601 [M+H]⁺.

Alternatively, the title compound was synthesized as described inExample 44 (Step 1-6) using 1-benzylpiperidine-4-carbaldehyde in step 6.¹H NMR (CDCl₃, 300 MHz) δ 7.27-7.33 (m, 5H), 6.89-6.96 (m, 4H), 4.66 (d,J=2.4 Hz, 2H), 4.11-4.16 (m, 1H), 4.00-4.05 (m, 1H), 3.80 (br, 2H), 3.69(br, 2H), 3.55 (br, 2H), 3.23-3.28 (m, 1H), 3.07-3.16 (m, 5H), 2.92 (br,2H), 2.79-2.82 (m, 1H), 2.52 (t, J=2.4 Hz, 1H), 2.28 (d, J=7.2 Hz, 2H),1.99-2.15 (m, 4H), 1.68-1.77 (m, 2H), 1.48-1.51 (m, 1H), 1.27-1.30 (m,2H). LCMS (ESI, m/z): 601 [M+H]⁺.

Example 47: Preparation of(R)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate (Compound 47)

7-((1-Benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylate (300 mg, 0.50 mmol,1.00 equiv) was separated by chiral-HPLC using the following gradientconditions: Column: Chiralpak IA 2*25 cm, 5 um; Mobile Phase A:acetonitrile (95%), Mobile Phase B: CH₂Cl₂ (5%); Flow rate: 20 mL/min;Detector, UV 254 nm; RT1: 11.50 min; RT2: 19.0 min. Chiral-HPLCseparation resulted in 59.5 mg (20% yield) of(R)-7-((1-benzylpiperidin-4-yl)methyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-(prop-2-ynyloxy)phenyl)piperazine-1-carboxylateas a white solid. ¹H NMR (CDCl₃, 300 MHz) δ 7.31-7.33 (m, 5H), 6.88-6.95(m, 4H), 4.65 (d, J=2.4 Hz, 2H), 4.10-4.15 (m, 1H), 3.99-4.04 (m, 1H),3.58-3.80 (m, 6H), 3.22-3.27 (m, 1H), 2.97-3.14 (m, 7H), 2.77-2.80 (m,1H), 2.51 (t, J=2.4 Hz, 1H), 2.28 (d, J=6.6 Hz, 2H), 2.04-2.14 (m, 4H),1.68-1.79 (m, 2H), 1.32-1.36 (m, 2H). LCMS (ESI, m/z): 601 [M+H]⁺.

Alternatively, the title compound was synthesized as described inExample 46 using (R)-tert-butyl2-hydroxy-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate asthe starting material. ¹H NMR (CDCl₃, 300 MHz) δ 7.27-7.34 (m, 5H),6.89-6.96 (m, 4H), 4.66 (d, J=2.4 Hz, 2H), 4.11-4.16 (m, 1H), 4.00-4.05(m, 1H), 3.80 (br, 2H), 3.68 (br, 2H), 3.56-3.57 (m, 2H), 3.23-3.28 (m,1H), 3.07-3.16 (m, 5H), 2.94 (br, 2H), 2.79-2.82 (m, 1H), 2.52 (t, J=2.4Hz, 1H), 2.28 (d, J=6.6 Hz, 2H), 2.06-2.15 (m, 4H), 1.69-1.78 (m, 2H),1.53 (br, 1H), 1.25-1.38 (m, 2H). LCMS (ESI, m/z): 601 [M+H]⁺.

Example 48: Preparation of(S)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylate (Compound 48)

7-(Morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylate(63.0 mg, 0.220 mmol, 1.00 equiv) was separated by Prep-Chiral-HPLCusing the following gradient conditions: Column: Chiralpak IA 2*25 cm, 5um; Mobile Phase: MeCN (100%); Flow rate: 20 mL/min; Detector, 254 nm;RT1: 8.5 min; RT2: 14.0 min. Chiral-HPLC separation resulted in 37.6 mg(59.7% yield) of(S)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylate as a white solid. ¹H NMR(CDCl₃, 300 MHz) δ 7.45 (d, J=9.0 Hz, 2H), 7.10 (d, J=9.0 Hz, 2H),4.10-4.28 (m, 2H), 4.02-4.09 (m, 3H), 3.66-3.71 (m, 5H), 3.30-3.34 (m,4H), 3.10-3.19 (m, 1H), 2.82-3.04 (m, 5H), 2.61-2.64 (m, 2H), 1.72-1.81(m, 2H), 1.56-1.63 (m, 2H), 1.40-1.52 (m, 1H), 1.30-1.40 (m, 2H). LCMS(ESI, m/z): 524 [M+H]⁺.

Alternatively, the title compound was prepared as follows.

Step 1: [(4-Iodophenyl)methyl]triphenylphosphanium bromide

A 250-mL round-bottom flask was charged with1-(bromomethyl)-4-iodobenzene (10.0 g, 33.7 mmol, 1.00 equiv, toluene(100 mL), triphenylphosphane (12.4 g, 47.3 mmol, 1.40 equiv). Theresulting solution was stirred overnight at 100° C. The solid wascollected by filtration to provide 20.0 g (crude) of[(4-iodophenyl)methyl]triphenylphosphanium bromide as a yellow solid.LCMS (ESI, m/z): 480 [M−Br]⁺

Step 2: tert-Butyl4-[(E)-2-(4-iodophenyl)ethenyl]piperidine-1-carboxylate

A 500-mL round-bottom flask was charged with[(4-iodophenyl)methyl]triphenylphosphanium bromide (20.0 g, 35.8 mmol,1.00 equiv), THF (250 mL), tert-butyl 4-formylpiperidine-1-carboxylate(7.62 g, 35.8 mmol, 1.00 equiv), and sodium hydride (60% w/w, 2.15 g,53.7 mmol, 1.50 equiv). The resulting solution was stirred for 5 h at 0°C. and quenched with water (300 mL). The mixture was extracted withEtOAc (3×100 mL) and the organic layers were combined, washed with brine(2×100 mL), dried over anhydrous Na₂SO₄, filtered and concentrated. Theresidue was chromatographed on a silica gel column with EtOAc/petroleumether (1/8) to provide 11.5 g (67% yield) of tert-butyl4-[(E)-2-(4-iodophenyl)ethenyl]piperidine-1-carboxylate as yellow oil.LCMS (ESI, m/z): 414 [M+H]⁺

Step 3: tert-Butyl 4-[2-(4-iodophenyl)ethyl]piperidine-1-carboxylate

A 50-mL round-bottom flask was charged with tert-butyl4-[(E)-2-(4-iodophenyl)ethenyl]piperidine-1-carboxylate (2.40 g, 5.81mmol, 1.00 equiv), ethylene glycol dimethyl ether (30 mL),4-methylbenzene-1-sulfonohydrazide (2.16 g, 11.6 mmol, 2.00 equiv),sodium acetate (1.43 g, 17.4 mmol, 3.00 equiv), water (8 mL). Theresulting solution was stirred overnight at 90° C. and quenched withwater (30 mL). The mixture was extracted with EtOAc (3×30 mL) and theorganic layers were combined, washed with brine (2×30 mL), dried overanhydrous Na₂SO₄, filtered and concentrated. The residue waschromatographed on a silica gel column with EtOAc/petroleum ether (1/10)to provide 1.60 g (66% yield) of tert-butyl4-[2-(4-iodophenyl)ethyl]piperidine-1-carboxylate as yellow oil. LCMS(ESI, m/z): 416 [M+H]⁺

Step 4: tert-Butyl4-(2-[4-[2-(trimethylsilyl)ethynyl]phenyl]ethyl)piperidine-1-carboxylate

A 50-mL round-bottom flask was charged with tert-butyl4-[2-(4-iodophenyl)ethyl]piperidine-1-carboxylate (2.40 g, 5.78 mmol,1.00 equiv), THF (30 mL), ethynyltrimethylsilane (850 mg, 8.65 mmol,1.50 equiv), copper (I) iodide (220 mg, 1.16 mmol, 0.20 equiv),bis(triphenylphosphine)palladium(II) chloride (406 mg, 0.578 mmol, 0.10equiv), and triethylamine (1.17 g, 11.6 mmol, 2.00 equiv). The resultingsolution was stirred overnight at 30° C. and quenched with water (10mL). The mixture was extracted with EtOAc (3×10 mL) and the organiclayers were combined, washed with brine (2×10 mL), dried over anhydrousNa₂SO₄, filtered and concentrated. The residue was chromatographed on asilica gel column with EtOAc/petroleum ether (1/10) to provide 2.00 g(90% yield) of tert-butyl4-[2-(4-[2-(trimethylsilyl)ethynyl]phenyl]ethyl)piperidine-1-carboxylateas a yellow solid. LCMS (ESI, m/z): 386 [M+H]⁺

Step 5: tert-Butyl 4-[2-(4-ethynylphenyl)ethyl]piperidine-1-carboxylate

A 50-mL round-bottom flask was charged with a solution of tert-butyl4-[2-(4-[2-(trimethylsilyl)ethynyl]phenyl]ethyl)piperidine-1-carboxylate(2.40 g, 6.22 mmol, 1.00 equiv) in THF (30 mL), and tetrabutylammoniumfluoride (2.44 g, 9.35 mmol, 1.50 equiv). The resulting solution wasstirred for 3 h at room temperature and quenched with water (10 mL). Themixture was extracted with CH₂Cl₂ (3×10 mL) and the organic layers werecombined, washed with brine (2×10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated. The residue was chromatographed on a silicagel column with EtOAc/petroleum ether (1/10) to provide 1.72 g (88%yield) of tert-butyl4-[2-(4-ethynylphenyl)ethyl]piperidine-1-carboxylate as yellow oil. LCMS(ESI, m/z): 314 [M+H]⁺

Step 6: 4-[2-(4-Ethynylphenyl)ethyl]piperidine

A 50-mL round-bottom flask was charged with tert-butyl4-[2-(4-ethynylphenyl)ethyl]piperidine-1-carboxylate (1.72 g, 5.49 mmol,1.00 equiv), CH₂Cl₂ (20 mL), and NMM (1.66 g, 16.5 mmol, 3.00 equiv).Trimethyiodosilane (2.20 g, 11.0 mmol, 2.00 equiv) was added dropwise.The resulting solution was stirred for 3 h at room temperature andquenched with water (10 mL). The mixture was extracted with CH₂Cl₂ (3×10mL) and the organic layers were combined, washed with brine (2×10 mL),dried over anhydrous Na₂SO₄, filtered and concentrated to provide 1.10 g(94% yield) of 4-[2-(4-ethynylphenyl)ethyl]piperidine as a yellow solid.LCMS (ESI, m/z): 214 [M+H]⁺

Step 7: 4-[2-(4-Ethynylphenyl)ethyl]piperidine-1-carbonyl chloride

A 50-mL round-bottom flask was charged with triphosgene (615 mg, 2.07mmol, 0.40 equiv) and CH₂Cl₂ (20 mL).4-[2-(4-Ethynylphenyl)ethyl]piperidine (1.10 g, 5.16 mmol, 1.00 equiv)was added. N,N-Diisopropylethylamine (2.00 g, 15.5 mmol, 3.00 equiv) wasadded dropwise. The resulting solution was stirred overnight at roomtemperature and quenched with water (10 mL). The mixture was extractedwith CH₂Cl₂ (3×10 mL) and the organic layers were combined, washed withbrine (2×10 mL), dried over anhydrous Na₂SO₄, filtered and concentratedto provide 1.40 g (98% yield) of4-[2-(4-ethynylphenyl)ethyl]piperidine-1-carbonyl chloride as brown oil.LCMS (ESI, m/z): 276 [M+H]⁺

Step 8:(S)-7-(Morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylate

A 50-mL round-bottom flask was charged with a solution of4-[2-(4-ethynylphenyl)ethyl]piperidine-1-carbonyl chloride (1.40 g, 5.08mmol, 1.00 equiv) in CH₂Cl₂ (20 mL),(S)-2-hydroxy-7-(morpholine-4-carbonyl)-hexahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-dione(1.74 g, 6.12 mmol, 1.21 equiv), NMM (1.55 g, 15.3 mmol, 3.00 equiv),and 4-DMAP (124 mg, 1.02 mmol, 0.20 equiv). The resulting solution wasstirred overnight at room temperature and quenched with water (10 mL).The mixture was extracted with CH₂Cl₂ (3×10 mL) and the organic layerswere combined, washed with brine (2×10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated. The crude product (400 mg) was purified bypreparative HPLC using the following gradient conditions: 30% CH₃CN/70%Phase A increasing to 70% CH₃CN over 10 min, then to 100% CH₃CN over 0.1min, holding at 100% CH₃CN for 1.9 min, then reducing to 30% CH₃CN over0.1 min, and holding at 30% for 1.9 min, on a Waters 2767-5Chromatograph. Column: Xbridge Prep C18, 19*150 mm 5 um; Mobile phase:Phase A: water; Phase B: CH₃CN; Detector, UV220 & 254 nm. Purificationresulted in 140 mg (5% yield) of(S)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylateas a white solid. ¹H NMR (CDCl₃, 300 MHz) δ 7.45 (d, J=9.0 Hz, 2H), 7.10(d, J=9.0 Hz, 2H), 4.10-4.28 (m, 2H), 4.02-4.09 (m, 3H), 3.66-3.71 (m,5H), 3.30-3.34 (m, 4H), 3.04-3.19 (m, 1H), 2.98 (s, 1H), 2.83-2.96 (m,4H), 2.61-2.67 (m, 2H), 1.77-1.81 (m, 2H), 1.56-1.63 (m, 2H), 1.40-1.49(m, 1H), 1.20-1.40 (m, 2H). LCMS (ESI, m/z): 524 [M+H]⁺.

Example 49: Preparation of(R)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylate (Compound 49)

7-(Morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylate(63.0 mg, 0.220 mmol, 1.00 equiv) was separated by Prep-Chiral-HPLCusing the following gradient conditions: Column: Chiralpak IA 2*25 cm, 5um; Mobile Phase: MeCN (100%); Flow rate: 20 mL/min; Detector, 254 nm;RT1: 8.5 min; RT2: 14.0 min. Chiral-HPLC separation resulted in 9.2 mg(14.6% yield) of(R)-7-(morpholine-4-carbonyl)-1,3-dioxo-hexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-ethynylphenethyl)piperidine-1-carboxylate as yellow solid. ¹H NMR(CDCl₃, 300 MHz) δ 7.45 (d, J=9.0 Hz, 2H), 7.10 (d, J=9.0 Hz, 2H),4.10-4.28 (m, 2H), 4.02-4.09 (m, 3H), 3.66-3.71 (m, 5H), 3.30-3.34 (m,4H), 3.04-3.19 (m, 1H), 2.98 (s, 1H), 2.83-2.96 (m, 4H), 2.61-2.67 (m,2H), 1.77-1.81 (m, 2H), 1.56-1.63 (m, 2H), 1.40-1.49 (m, 1H), 1.20-1.40(m, 2H). LCMS (ESI, m/z): 524 [M+H]⁺.

Example 50: Preparation of(S)-7-(morpholine-4-carbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chloro-phenethyl)piperidine-1-carboxylate (Compound 50)

The title compound was separated from7-(morpholine-4-carbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chloro-phenethyl)piperidine-1-carboxylate (Example 1) following theprocedure described in Example 40.

Example 51: Preparation of(R)-7-(morpholine-4-carbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chloro-phenethyl)piperidine-1-carboxylate (Compound 51)

The title compound was separated from7-(morpholine-4-carbonyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl4-(4-chloro-phenethyl)piperidine-1-carboxylate (Example 1) following theprocedure described in Example 40.

II. Biological Evaluation

Assays:

Transient Overexpression

HEK293T cells were seeded at 2×105 per well on 6 well plates and grownfor 48 h. They were then transfected with mABHD3 (pCMV-Sport6), hABHD4(pcDNA3.1/myc-his), hPPT1 (pCMV6-XL5), or hPPT1 S115A (pCMV6-XL5), byincubating 0.5 μg of DNA with 3 μL of PEI MAX (1 mg/mL, Polysciences,Inc) in serum-free DMEM for 30 min, then adding them to cells. After 48h cells were either treated with compound in situ, as described above,or harvested for in vitro studies. The S110A PPT1 mutation wasintroduced using QuikChange II Site Directed Mutagenesis Kit (Agilent)with the following primers:

hPPT1 S→A QC Forward: aattggcctccctgggcgaatcccatagcattghPPT1 S→A QC Reverse: caatgctatgggattcgcccagggaggccaattWestern Blotting

Following SDS-PAGE, PPT1 samples were transferred to nitrocellulose for2 h at 50 V. Transfers were then washed, blocked with 5% milk, andincubated overnight with Anti-PPT1 (Abcam ab89022, 1:2000 in TBS-T with5% milk). Transfers were then washed 3× (TBS-T) and incubated with asecondary antibody (Odyssey 926-23310, 1:5000 in TBS-T with 5% milk) for2 h. Transfers were then washed again (3×, TBS-T) and imaged on a Li-corOdyssey (Model 9120).

PPT1 Substrate Assay

PPT1 activity was assessed in vitro using a substrate assay adapted fromprevious studies (van Diggelen et al., Molecular genetics and metabolism66, 240-244, 1999). Lysates from HEK293T cells transfected with an emptyvector, or human PPT1 under a CMV promoter were adjusted to 1 mg/ml inPBS (25 μl), and then combined 1:1 with Mcllvain's phosphate/citratebuffer containing 0.375% (v/v) Triton-X 100 and 15 mM DTT (pH 5.0). Thissample was pre-treated with inhibitor or DMSO for 30 minutes at 37° C.,and 10 μl (5 μg protein) was used in the substrate assay as follows:Samples were diluted in a black 96-well plate (half-area) with 20 μlMcIlvain's phosphate/citrate buffer (pH 5.0) containing 0.375% TX-100and 15 mM DTT, as well as 0.64 mM MU-6S-Palm-PD-Glc and 0.1 U sweetalmond glucosidase (Sigma). The reactions were incubated at 37° C. for 1hour, quenched with 120 μl sodium bicarbonate buffer (pH 10.0), andhydrolysis by 4-MU fluorescence measured by excitement at 380 nM andemission intensity at 460 nM. 16 μg protein was used for analysis ofnative PC3 cells.

IsoTOP Site-of-Labeling of PPT1

Whole cell lysates (500 μL, 1.5 mg/mL) of hPPT1 transfected HEK239Tswere processed for MS analysis using the previously describedisoTOP-ABPP protocol (Weerapana et al., Nature 468, 790-795, 2010). Inbrief, for analysis of Compound 39-modified peptides, proteomes weresplit into two fractions, to which Compound 39 (5 μM, 60 min) or DMSOwere added. The lysates were then subjected to Click Chemistry-ABPPconditions with either light (Compound 39 treated samples) or heavy(DMSO treated samples) isotopically labeled TEV-tags. Light andheavy-tagged proteomes were then combined, and, following enrichment ofprobe-labeled targets using streptavidin beads, proteins were digestedon-bead with trypsin and remaining immobilized peptides released with asubsequent TEV protease digestion. The resulting probe-modified peptideswere pressure loaded onto a 100 μm (inner diameter) fused silicacapillary column with a 5 μm tip containing 10 cm C18 resin (5 μm,Phenomenex), eluted with a 180 min gradient from 0% to 100% Buffer B(Buffer A: 5% acetonitrile, 95% water, 0.1% formic acid; Buffer B: 80%acetonitrile, 20% water, 0.1% formic acid), and collected for MSanalysis on an LTQ-Orbitrap. Samples were then searched as describedabove. The mass of the modification used to search for probe-modifiedpeptides was +626.3289 m/z for the probe adduct plus the light TEV-tagand +632.3427 m/z for the heavy counterpart. MS1 peaks for probe-labeledquantified peptides were extracted with Xcalibur Qual Browser 2.2(Thermo Scientific).

Example A1: Solubility and Stability Assays

Solubility in PBS:

The solubility of compounds were tested in triplicate in phosphatebuffered saline (PBS), pH 7.4. Per well, 198 μL PBS is added to aMillipore Solvinert Hydrophilic PTFE 96 well filter plate: pore size:0.45 μm (MSRLN0450). Test compounds were introduced from 10 mM DMSOstock solutions (2 μL). The final concentration of DMSO was 1 percent.Samples were allowed to incubate at 22° C. for 18 hours. In the morningthe plate was centrifuged where the soluble portion passes through thefilter and was collected in a capture plate. Clotrimazole was includedas a control to assure the assay was working properly. The samples wereanalyzed by HPLC. Peak area was compared to a standard of knownconcentration. In cases when the concentration was too low for UVanalysis or when the compound did not possess a good chromophore,LC-MS-MS analysis was used.

The solubility of Compound 1 was determined to be 18 μM in PBS.

Solubility in Media: The solubility of compounds were tested intriplicate in complete media (DMEM+10% FBS). Per well, 198 μL PBS isadded to a Millipore Solvinert Hydrophilic PTFE 96 well filter plate:pore size: 0.45 μm (MSRLN0450). Test compounds were introduced from 10mM DMSO stock solutions (2 μL). The final concentration of DMSO was 1percent. Samples were allowed to incubate at 22° C. for 18 hours. In themorning the plate was centrifuged where the soluble portion passedthrough the filter and was collected in a capture plate. The sampleswere analyzed by HPLC (Agilent 1100 with diode-array detector). Peakarea was compared to a standard of known concentration. In cases whenthe concentration was too low for UV analysis or when the compound didnot possess a good chromophore, LC-MS-MS analysis was used.

The solubility of compound 1 was determined to be 46 μM in DMEMcontaining 10% fecal calf serum.

Stability in PBS:

Demonstration of stability in PBS was conducted by addition of 10 μMcompound from a DMSO stock to PBS in HPLC autosampler vials. Sampleswere held in the HPLC autosampler at ambient temperature. Atapproximately 0, 1, 2, 4, 8, 24, and 48 hours the samples were injectedon the HPLC. Peak area and retention time were compared betweeninjections. Data was log transformed and represented as half-life. DMSOwas added as a co-solvent as needed for solubility.

The stability of compound 1 was determined to be 19 h in PBS, likely dueto hydrolytic opening of the NHS group.

Example A2: Determination of Glutathione Reactivity

Compound (10 μM) was incubated at 37° C. for 6 hours in the presence of50 μM freshly prepared reduced glutathione. At 0 and 6 hours the sampleswere injected into the HPLC instrument. Peak area and retention timewere compared between injections. Samples were evaluated for aglutathione dependent decrease in compound concentration. DMSO was addedas a co-solvent as needed for solubility.

Compound 1 did not show any reactivity with glutathione (50 μM),indicating that the compound has a tempered electrophilicity andspecific structural elements that direct reactivity towards a highlyselective enzyme subset (LYPLAs and ABHD6). Gel-based and LC-MS/MS(MudPIT)-based ABPP selectivity profiling also confirmed high targetspecificity for over 20 SHs.

Example A3: Gel-Based ABPP Analysis of Potency and Selectivity In Vitro

Assay Overview: The purpose of this assay was to determine whether testcompounds can inhibit LYPLA1 and LYPLA2 in a complex proteomic lysateand to assess anti-target inhibition using a competitive activity-basedproteomic profiling (ABPP) assay. In this assay, a complex proteomeendogenously expressing LYPLA1/2 was incubated with test compoundfollowed by reaction with a rhodamine-conjugated fluorophosphonate(FP-Rh) serine-hydrolase specific activity-based probe. The reactionproducts were separated by SDS-PAGE and visualized in-gel using aflatbed fluorescence scanner. The percentage activity remaining wasdetermined by measuring the integrated optical density of the bands. Asdesigned, test compounds that act as LYPLA1 and/or LYPLA2 inhibitorsprevented enzyme-probe interactions, thereby decreasing the proportionof bound fluorescent probe, giving lower fluorescence intensity in theband in the gel. Percent inhibition was calculated relative to a DMSO(no compound) control.

Protocol Summary: Mouse brain membrane proteome (50 μL reaction volume,1 mg/mL in DPBS) was treated with 0.1-100 μM test compound (1 μL of a50× stock in DMSO) for 30 minutes at 37° C. FP-Rh (1 μL of 50× stock inDMSO, Thermo #88318) was added to a final concentration of 2 μM. Thereactions were incubated for 30 minutes at 25° C., quenched with 16 μL4×SDS-PAGE loading buffer (reducing), separated by SDS-PAGE andvisualized by in-gel fluorescent scanning. The percentage activityremaining was determined by measuring the integrated optical density ofthe bands relative to a DMSO-only (no compound) control. Assay Cutoff:Compounds with ≥50% inhibition of targets at 1 μM test compoundconcentration were considered active.

N-Hydroxy bicyclic hydantoin carbamates (Compounds 1-20) listed in Table1 were subjected to gel-based competitive ABPP screening to assess SHreactivity against more than 20 FP-sensitive SHs visible by 1D SDS-PAGEseparation and fluorescent detection in the mouse brain proteome (seeFIG. 1). Mouse brain was selected due to its abundance of SH enzymes andhigh homology between human and mouse SHs (e.g., comparable potency formouse and human isoforms of LYPLAs observed for Compound 1). Compoundtreatment (0.1-1000 nM, 30 minutes) was followed by labeling withSH-specific ABPP probe FP-Rh (2 μM, 30 minutes), 1D-SDS-PAGE, and in-gelfluorescent visualization of FP-labeled proteins. Assignment of LYPLA1/2bands was based on correlation of molecular weights, expression andinhibition profiles with LC-MS/MS data and recombinant enzymeexpression. Proteins were listed as anti-targets if at least 50%inhibition was observed as quantified relative to the DMSO control.Compound 1 exhibited the best combination of potent target inhibitionand high selectivity; other than ABHD6, out of the 20+ SHs visible bygel, the only anti-target with appreciable inhibition at a dose below 10μM was FAAH (IC₅₀=3150 nM, selectivity of ˜13-fold).

Compounds 1-20 were subjected to gel-based competitive ABPP profiling toassess potency and selectivity against several dozen FP-sensitive SHs inthe mouse brain membrane proteome, which endogenously expresses LYPLA1and LYPLA2, as well a diversity of potential SH anti-targets. Compoundswere tested at 100 μM, 10 μM, 1 μM, and/or 0.1 μM compoundconcentration.

Tables 2 and 3 summarize IC₅₀ and % inhibition as well as anti-targetactivity for Compounds 1-20.

TABLE 2 LYPLA1 LYPLA2 % INH^(‡) % INH^(‡) Anti-target(s)^(¶) Cpd. IC₅₀^(§) 100 μM 10 μM 1 μM 0.1 μM IC₅₀ ^(§) 100 μM 10 μM 1 μM 0.1 μM 100 μM10 μM 1 μM 0.1 μM 1 122 100 100 100 NT* 245 100 100 100 NT FAAH, FAAH,ABHD6 NT ABHD12, ABHD6 MAGL, ABHD6 2 154 100 100 100 50 258 100 100 10050 FAAH, FAAH, FAAH, ABHD6 ABHD12, ABHD12, ABHD6 ABHD4, ABHD4, MAGL,MAGL, ABHD6 ABHD6 225 100 100 100 50 384 100 100 100 50 FAAH, FAAH,FAAH, ABHD6 ABHD12, ABHD6 ABHD6 ABHD4, MAGL, ABHD6 4 92 100 100 100 80217 100 100 100 80 FAAH, FAAH, FAAH, ABHD6 ABHD12, ABHD12, ABHD6 ABHD4,MAGL, MAGL, ABHD6 ABHD6 5 NT 100 100 80 50 NT 100 100 80 50 FAAH, FAAH,FAAH, ABHD6 ABHD12, ABHD12, ABHD6 MAGL, ABHD6 ABHD6 6 NT 100 100 95 25NT 100 100 95 25 FAAH, FAAH, FAAH, ABHD6 MAGL, MAGL, ABHD6 ABHD6 ABHD6 7306 100 95 80 0 726 100 95 80 0 FAAH, FAAH, ABHD6 ABHD6 ABHD12, ABHD12,ABHD4, ABHD6 MAGL, ABHD6 8 NT 100 100 75 NT NT 100 100 75 NT FAAH, FAAH,ABHD6 NT ABHD12, ABHD12, MAGL, ABHD6 ABHD6 9 NT 100 100 90 NT NT 100 10090 NT FAAH, FAAH, ABHD6 NT ABHD12, ABHD12, MAGL, ABHD6 ABHD6 10 NT 100100 75 NT NT 100 100 75 NT FAAH, FAAH, ABHD6 NT ABHD12, ABHD12, ABHD6ABHD6 11 NT 100 100 90 NT NT 100 100 90 NT FAAH, FAAH, ABHD6 NT ABHD12,ABHD12, MAGL, MAGL, ABHD6 ABHD6 12 NT 100 95 25 0 NT 100 95 25 0 FAAH,FAAH, ABHD6 ABHD6 ABHD4, ABHD4, MAGL, MAGL, ABHD6 ABHD6 13 NT 90 75 10 0NT 90 75 10 0 FAAH, FAAH, FAAH, ABHD6 ABHD4, ABHD4, ABHD4, MAGL, ABHD6ABHD6 ABHD6 ^(§)IC₅₀ values were determined from three replicates ateach inhibitor concentration against the human LYPLA isoforms(comparable values obtained for mouse isoform) ^(‡)% Inhibition valueswere determined from one replicate at each inhibitor concentration *NT =not tested ^(¶)SHs are listed as anti-targets if at least 50% inhibitionwas observed at a given compound concentration (see Table 7 for enzymenames)

TABLE 3 LYPLA1 LYPLA2 % INH^(‡) % INH^(‡) Anti-target(s)^(¶) Cpd. 100 μM10 μM 1 μM 0.1 μM 100 μM 10 μM 1 μM 0.1 μM 100 μM 10 μM 1 μM 0.1 μM 1425 0 0 0 25 0 0 0 APEH, ABHD6 ABHD6 ABHD6 ABHD6 15 25 0 0 0 25 0 0 0ABHD6 ABHD6 ABHD6 ABHD6 16 0 0 0 0 0 0 0 0 ABHD6 ABHD6 none none 17 9075 0 0 75 50 0 0 FAAH, ABHD6 ABHD6 ABHD6 ABHD12, ABHD6 18 60 50 10 0 6050 10 0 ABHD6 ABHD6 ABHD6 ABHD6 19 50 10 0 0 25 0 0 0 FAAH, ABHD6 ABHD6ABHD6 ABHD6 20 100 100 10 0 100 75 10 0 ABHD6 ABHD6 ABHD6 ABHD6 ^(‡)%Inhibition values were determined from one replicate at each inhibitorconcentration ^(¶)SHs are listed as anti-targets if at least 50%inhibition was observed at a given compound concentration (see table 7for enzyme names)

In vitro enzyme activities (mouse brain (mem) proteome) for Compounds21, 23, 31, 38, 40-53 are shown in Table 4.

TABLE 4 Cpd. ABHD3 FAAH ABHD12 ABHD6 PLA2G7 LYPLA1 LYPLA2 21 100% @ 0% @0% @ 20 μM 20 μM 20 μM 23 100% @ 0% @ 0% @ 20 μM 20 μM 20 μM 31 0% @ 0%@ 20 μM 20 μM 38 100% @ 0% @ 0% @ 20 μM 20 μM 20 μM 40 0% @ 50% @ 0% @100% @ 100% @ 50% @ 50% @ 10 μM 10 μM 10 μM 1 μM 10 μM * 10 μM 10 μM 410% @ 100% @ 0% @ 100% @ 0% @ 100% @ 100% @ 10 μM 10 μM 10 μM 1 μM 10 μM10 μM 10 μM 42 0% @ 0% @ 0% @ 100% @ 100% @ 50% @ 100% @ 10 μM 10 μM 10μM 1 μM 1 μM 10 μM 10 μM 43 0% @ 0% @ 0% @ 100% @ 100% @ 0% @ 0% @ 10 μM10 μM 10 μM 1 μM 1 μM 10 μM 10 μM 44 0% @ 0% @ 0% @ 100% @ 100% @ 0% @0% @ 10 μM 10 μM 10 μM 1 μM 1 μM 10 μM 10 μM 45 75% @ 0% @ 0% @ 100% @100% @ 0% @ 0% @ 10 μM 10 μM 10 μM 1 μM 1 μM 10 μM 10 μM 46 0% @ 0% @ 0%@ 100% @ 100% @ 50% @ 50% @ 10 μM 10 μM 10 μM 1 μM 1 μM 10 μM 10 μM 470% @ 0% @ 0% @ 100% @ 100% @ 0% @ 0% @ 10 μM 10 μM 10 μM 1 μM 1 μM 10 μM10 μM 48 0% @ 75% @ 50% @ 100% @ 75% @ 75% @ 75% @ 10 μM 10 μM 10 μM 1μM 10 μM 1 μM 1 μM 49 0% @ 50% @ 0% @ 100% @ 100% @ 75% @ 75% @ 10 μM 10μM 10 μM 1 μM 10 μM 1 μM 1 μM 50 0% @ 0% @ 0% @ 100% @ 100% @ 100% @100% @ 10 μM 10 μM 10 μM 1 μM 10 μM 1 μM 1 μM 51 0% @ 0% @ 0% @ 100% @100% @ 100% @ 100% @ 10 μM 10 μM 10 μM 1 μM 10 μM 1 μM 1 μM 52 0% @ 0% @0% @ 100% @ 50% @ 0% @ 0% @ 10 μM 10 μM 10 μM 1 μM 1 μM 10 μM 10 μM 530% @ 50% @ 50% @ 100% @ 0% @ 75% @ 75% @ 10 μM 10 μM 10 μM 1 μM 10 μM 1μM 1 μM

In vitro potencies (IC₅₀, nM) for Compounds 31, 34-38, 42, 48-51 areshown in Table 5.

TABLE 5 Cpd. mLYPLA1/2 mABHD6 hPPT1 mABHD3 hABHD4 hLYPLA1/2 31 >10,0008200 34 >10,000 ~1000 35 >10,000 1000 36 >10,000 200 37 7600 100 38 10030 42 29300 3971 48 1750/ 1829 49 842/ 1434 50 117 2.2 51 97 2.5

Table 6 shows in situ and in vivo enzyme activity data for Compounds24-26, 34, and 37.

TABLE 6 hLYPLA1/2 mLYPLA1/2 Cpd. hPPT1 (in situ) mPPT1 (in vivo) (insitu) (in vivo) 1 500 nM 100% @ 10 mg/kg 87/143 nM 100% @ (testis) 5mg/kg (liver and kidney) 24 100% @ 2 μM 25 100% @ 2 μM 26 100 nM 100% @5 mg/kg 0% @ (testis) 40 mg/kg (testis) 34 3900 nM 37 100% @ 2 μM

Table 7 shows the enzyme names and identifiers.

TABLE 7 Assay Target Name GeneID Protein GI Taxonomy ABHD4 alpha/betahydrolase 105501 326937491 Mus domain containing musculus protein 4ABHD6 alpha/beta hydrolase 66082 31560264 Mus domain containing musculusprotein 6 57406 189027141 Homo sapiens ABHD10 alpha/beta hydrolase 553478923001 Homo domain containing sapiens protein 10 ABHD11 alpha/betahydrolase 83451 74751292 Homo domain-containing sapiens protein 11ABHD12 alpha/beta hydrolase 26090 109689718 Homo domain containingsapiens protein 12 76192 159110817 Mus musculus ACOT2 acyl-coenzyme A10965 269849771 Homo thioesterase 2 sapiens APEH acylpeptide hydrolase327 23510451 Homo sapiens 235606 19343726 Mus musculus CTSA lysosomalprotective 5476 20178316 Homo protein sapiens DPP8 dipeptidyl peptidase8 54878 67460301 Homo sapiens ESD esterase 2098 33413400 HomoD/formylglutathione sapiens hydrolase FAAH fatty acid amide 14073123253900 Mus hydrolase musculus FAM108B1 alpha/beta hydrolase 5110471051600 Homo domain-containing sapiens protein 17B isoform 1 precursorFAP isoform 1 of Seprase 2191 292495099 Homo sapiens LYPLA1lysophospholipase 1 10434 5453722 Homo sapiens 18777 71059731 Musmusculus LYPLA2 lysophospholipase 2 11313 9966764 Homo sapiens 26394123122209 Mus musculus LYPLAL1 lysophospholipase- 127018 20270341 Homolike 1 sapiens MAGL monoacylglycerol 11343 6005786 Homo (MGLL) lipasesapiens 23945 261878511 Mus musculus NCEH1 neutral cholesterol 5755268051721 Homo ester hydrolase 1 sapiens PAFAH1B2 platelet-activating5049 55977294 Homo factor sapiens acetylhydrolase IB subunit betaPAFAH1B3 platelet-activating 5050 3024344 Homo factor sapiensacetylhydrolase IB subunit gamma PAFAH2 platelet-activating 5051 6647691Homo factor sapiens acetylhydrolase 2 PLA2G15 group XV 23659 44888104Homo phospholipase A2 sapiens PNPLA4 patatin-like 8228 116242718 Homophospholipase domain- sapiens containing protein 4 PNPLA6 patatin-like10908 260656037 Homo phospholipase domain sapiens containing protein 6PRCP lysosomal Pro-X 5547 1172047 Homo carboxypeptidase sapiensprecursor PREPL prolyl endopeptidase- 9581 121944206 Homo like sapiensSCPEP1 isoform 1 of Retinoid- 59342 11055992 Homo inducible serinesapiens carboxypeptidase precursor

Example A4: Determination of IC₅₀ by Gel-Based Competitive ABPP In Vitro

Assay Overview: The purpose of this assay was to determine IC₅₀ valuesof powder samples of test compounds for enzyme of interest activity, andanti-targets in a complex proteomic lysate using a gel-based competitiveABPP assay. In this assay, a complex proteome was incubated with testcompound followed by reaction with FP-Rh and SDS-PAGE analysis asdescribed above.

Protocol Summary: Soluble HeLa cell proteome (50 μL reaction volume, 1mg/mL in DPBS) or mouse brain membrane proteome (50 μL reaction volume,1 mg/mL in DPBS) was treated with varying concentrations of testcompound (1 μL of a 50× stock in DMSO) for 30 minutes at 37° C. FP-Rh (1μL of 50× stock in DMSO) was added to a final concentration of 2 μM. Thereaction was incubated for 30 minutes at 25° C., quenched with 16 μL4×SDS-PAGE loading buffer (reducing), separated by SDS-PAGE andvisualized by in-gel fluorescent scanning. The percentage activityremaining was determined by measuring the integrated optical density ofthe enzymes of interest, and anti-target bands relative to a DMSO-only(no compound) control. IC₅₀ values were determined from dose-responsecurves from three replicates at each inhibitor concentration: 1, 10,100, 1000, 10000 nM and 0.01, 0.1, 1, 10, 100, 1000, 10000, 50000, and100000 nM. Assay Cutoff: Compounds with an IC₅₀ less than or equal to1000 nM were considered active.

Example A5: In Situ Inhibitor Treatment of Human Cell Lines

In situ inhibitor treatments of human cell lines were performed asdescribed previously (Hsu et al., J Med Chem 56, 8270-8279, 2012).Briefly, PC3 or transfected HEK293T cells were grown to confluency inserum-containing media (RPMI or DMEM, respectively). The culture mediawas then removed and replaced with fresh media containing DMSO orinhibitor at the desired concentration and incubated for 4 hours in atissue culture incubator (37° C., 5% CO₂). Cells were then washed withPBS, harvested, washed with PBS once more, and lysed with a probesonicator.

Example A6: Determination of IC₅₀ Values by Gel-Based Competitive ABPPIn Situ

Assay Overview: The purpose of this assay was to determine IC₅₀ valuesof powder samples of test compounds for enzyme of interest activity insitu using a gel-based competitive ABPP assay. In this assay, culturedcells were treated with test compound, harvested, and lysed. Theisolated soluble proteome fraction was reacted with FP-Rh and analyzedby SDS-PAGE as described above.

Protocol Summary: To cultured HeLa cells (90% confluent) was added freshDMEM (5 mL total volume; supplemented with 10% FCS) pre-mixed with DMSOor test compound. After 2 hours at 37° C., cells were harvested byscraping, washed twice with 10 mL DPBS, and homogenized by sonication inDPBS. The soluble fraction was isolated by centrifugation (100K×g, 45minutes) and the protein concentration was adjusted to 1 mg/mL withDPBS. FP-Rh (1 μL of 50× stock in DMSO) was added to a finalconcentration of 2 μM in 50 μL total reaction volume. The reaction wasincubated for 30 minutes at 25° C., quenched with 16 μL 4×SDS-PAGEloading buffer (reducing), separated by SDS-PAGE and visualized byin-gel fluorescent scanning. The percentage activity remaining wasdetermined by measuring the integrated optical density of the enzymes ofinterest, and anti-target bands relative to a DMSO-only (no compound)control. IC₅₀ values were determined from dose-response curves fromthree replicates at each inhibitor concentration: 0.1, 1, 50, 250, 1000,20000 nM. Assay Cutoff: Compounds with an IC₅₀ less than or equal to1000 nM were considered active.

IC₅₀ values for Compound 1 against the human isoforms of LYPLA1 (122 nMin vitro and 87 nM in situ) and LYPLA2 (245 nM in vitro and 143 nM insitu) were obtained from gel-based competitive-ABPP data using the FP-Rhactivity-based probe in both HeLa cell lysates (see FIG. 2A) andcultured HeLa cells (see FIG. 2B). Compound 1 shows comparable in vitroinhibition (collective IC₅₀=154 nM) of the mouse isoforms of LYPLA1/2(see FIG. 2C; note: LYPLA1/2 quantified together due to the overlappingnature of the target bands). IC₅₀ values were also determined foranti-targets ABHD6 and FAAH (see FIG. 2C), with Compound 1 showingpotent (IC₅₀=3.15 nM) inhibition of ABHD6, but good selectivity for FAAH(IC₅₀=3150 nM, ˜13-fold selectivity).

Compound 1 is active in situ against the LYPLA enzymes (see FIG. 3A) asassessed by gel-based competitive ABPP with FP-Rh following two hours ofcompound treatment of HeLa cells cultured in serum-supplemented medium.(IC₅₀ values of 87 nM and 143 nM were calculated for LYPLA1 and LYPLA2,respectively). No anti-target inhibition is observed at any compoundconcentration (ABHD6 not visible).

Example A7: Inhibition by Gel-Based Competitive ABPP In Vivo

Assay Overview: The purpose of this assay was to determine whether ornot powder samples of test compounds inhibit enzymes of interest invivo. In this assay, test compounds were administered to mice. Mice weresacrificed, and their tissues harvested, homogenized, and the solublefractions isolated, reacted with FP-Rh, and separated by SDS-PAGE asdescribed above.

Protocol Summary: Purpose-bred 3-4 month old male C57BL6 laboratory micewere i.p. administered test compound (0.5-50 mg/kg in 18:1:1saline:PEG-40-castor oil:EtOH vehicle solution, 10 μL/g mouse weight) orvehicle only. After 4 hours, mice were humanely sacrificed (anesthetizedwith isoflurane followed by cervical dislocation), and tissues (liver,kidney, brain) removed and snap frozen in liquid nitrogen beforeprocessing. Tissues were homogenized, and soluble proteome isolated byultracentrifugation (100 k×g, 45 minutes) and protein concentrationadjusted to 1 mg/mL in DPBS. An aliquot (50 μL) was reacted with FP-Rh(2 μM final concentration) for 30 minutes at 25° C. Reactions werequenched with 16 μL 4×SDS-PAGE loading buffer (reducing), separated bySDS-PAGE, and visualized by in-gel fluorescent scanning. The percentageactivity remaining was determined by measuring the integrated opticaldensity of test compound bands relative to vehicle bands. Note: Due tothe overlapping nature of LYPLA1 and LYPLA2 bands, the proteins werequantified together. Assay Cutoff: Compounds with greater than or equalto 50% inhibition at 5 mg/kg test compound concentration in one or moretissues tested were considered active.

The in vivo inhibitory activity of Compound 1 was assessed. For thisexperiment, mice were administered test compound (0.5-50 mg/kg, i.p.) orvehicle only. After four hours mice were sacrificed and their tissuesremoved, homogenized, and the soluble fractions isolated and labeledwith FP-Rh followed by SDS-PAGE and fluorescent detection of FP-labeledSHs (see FIG. 3B).

Near-complete (˜90%) inhibition of LYPLA1/2 was observed in liver andkidney. In contrast, only ˜75% inhibition was observed in the brain atthe highest dose (50 mg/kg). Among the 20+ distinct SH bands, nosignificant anti-target inhibition was observed in any tissue (ABHD6 notvisible); these data indicate that Compound 1 is active and selective invivo, and that target inhibition appears largely restricted toperipheral tissues.

Example A8: Selectivity Analysis by ABPP-SILAC

Competitive ABPP-SILAC (Adibekian, A., et al. J Am Chem Soc, 2012.134(25): p. 10345-8) combined competitive ABPP (Leung, D., et al. NatBiotechnol, 2003. 21(6): p. 687-91) with stable isotope labeling ofcells (SILAC) (Ong, S. E., et al. Mol Cell Proteomics, 2002. 1(5): p.376-86), and allowed for precise quantitation of enzyme inhibition bycalculating the isotopic ratios of peptides from inhibitor-treated andcontrol cells.

Assay Overview: The purpose of this assay was to determine theselectivity profile of powder samples of test compounds using stableisotope labeling with amino acids in cell culture (SILAC) ABPP. In thisassay, cultured HeLa cells were metabolically labeled with light orheavy amino acids. Light and heavy cells were treated with inhibitor andDMSO, respectively, in situ. Cells were harvested, and proteomefractions were isolated and reacted with the serine-hydrolase-specificactivity-based affinity probe fluorophosphonate-biotin (FP-biotin).Light and heavy samples were combined in a 1:1 (w/w) ratio. Biotinylatedproteins were enriched, trypsinized, and analyzed by LC/LC-MS/MS(MudPIT) Inhibition of target and anti-target activity was quantified bycomparing intensities of light and heavy peptide peaks. As designed,compounds that acted as inhibitors blocked FP-biotin probe labeling,reducing enrichment in the inhibitor-treated (light) sample relative tothe DMSO-treated (heavy) sample, giving a smaller light/heavy ratio foreach protein. Proteins not targeted by inhibitors were expected to havea ratio close to 1.

Sample Preparation. HeLa cells were initially grown for at least eightpassages in either light or heavy SILAC DMEM medium supplemented with10% dialyzed FCS and 1× PenStrep Glutamine in a humidified incubator(37° C., 5% CO2). Light medium was supplemented with 100 μg/mLL-arginine (Sigma) and 100 μg/mL L-lysine (Sigma). Heavy medium wassupplemented with 100 μg/mL [13C615N4]-L-Arginine (Isotek) and 100 μg/mL[13C615N2]-L-Lysine (Isotek). Light cells were treated with testcompound (1 μM, from a 1000× stock in DMSO) and heavy cells were treatedwith DMSO for 2 hours at 37° C. Cells were washed once with cold DPBS,harvested by scraping, and the quadruplicate samples for each conditionpooled and lysed in DPBS by sonication. The pooled whole-cell extractswere adjusted to 2 mg/mL [protein], and treated with 10 μM FP-biotin for1 hour at room temperature in 500 μL DPBS (1 mg protein per condition).Light and heavy samples were then combined 1:1 (w/w protein), and thetotal protein was precipitated from the samples by addition of 1 mL MeOHand 250 μL CHCl₃, followed by vortexing. The protein was pelleted bycentrifugation (1400×g, 20 minutes, 4° C.), the supernatant discarded,and the pellet then washed twice more with 1:1 MeOH/CHCl₃ (500 μL, then250 μL, with the protein pellet resuspended by sonication and pelletedby centrifugation, 16 k×g for 10 minutes, following each wash). Thewashed protein pellets were solubilized first by sonication in 6M ureain aqueous 25 mM ammonium bicarbonate, and then by addition of 140 μL10% aqueous SDS. Samples were reduced with 10 mM DTT at 65° C. for 15minutes, and then alkylated with 10 mM iodoacetamide for 30 minutes atroom temperature in the dark. Samples were added directly to 6 mL DPBScontaining 50 μL avidin-agarose, and biotinylated proteins were thenenriched from each sample by incubation over the avidin beads for 1.5hours with agitation at room temperature. The beads were washed oncewith 1% SDS, then three times with DPBS, and then transferred toscrew-cap tubes in 200 μL 2 M urea/25 mM ammonium bicarbonate. Calciumchloride was added to 2 mM final, and on-bead digestion was performedfor 12 hours at 37° C. with sequencing grade modified trypsin (2 μg;Promega). Beads were then filtered from each sample and the resultantpeptide samples were acidified with 16 μL formic acid (5% v/v) andstored at −80° C. until analysis.

LC-MS/MS analysis. Samples were analyzed by multidimensional liquidchromatography tandem mass spectrometry (MudPIT) using an Agilent1200-series quaternary pump and Thermo Scientific LTQ Orbitrap Velos iontrap mass spectrometer. Peptides were eluted in a 5-step MudPITexperiment using 0%, 25%, 50%, 80%, and 100% salt bumps of 500 mMaqueous ammonium acetate and data were collected in data-dependentacquisition mode with dynamic exclusion turned on (20 seconds, repeatcount of 1). Specifically, one full MS (MS1) scan (400-1800 m/z) wasfollowed by 30 MS2 scans of the most abundant ions. The MS2 spectra datawere extracted from the raw file using RAW Xtractor (version 1.9.9.2;publicly available at http://fields.scripps.edu/researchtools.php). MS2spectra data were searched using the Sequest algorithm against thelatest version of the human UNIPROT database concatenated with thereversed database for assessment of false-discovery rates. Sequestsearches allowed for static modification of cysteine residues (+57.02146due to alkylation), methionine oxidation (+15.9949), mass shifts oflabeled amino acids (+10.0083 R, +8.0142 K) and no enzyme specificity.The resulting MS2 spectra matches were assembled into proteinidentifications and filtered using DTASelect (version 2.0.41; publiclyavailable at http://fields.scripps.edu/researchtools.php) using the—trypstat option (applies different statistical models for the analysisof peptide digestion state), and a maximum false-positive rate set to1%. Ratios of light/heavy (cpd/DMSO) peaks were calculated usingin-house software and normalized at the peptide level to the averageratio of all non-serine hydrolase peptides.

Reported ratios represent the mean of all unique, quantified peptidesper protein and do not include peptides that were >3 standard deviationsfrom the median peptide value. Proteins with less than three peptidesper protein ID were not included in the analysis. Assay Cutoff Acompound was considered active for a particular target/anti-target witha light/heavy ratio of ≤0.5.

The results (see FIG. 4A) indicated that Compound 1 achievednear-complete (>90%) inhibition of human LYPLA1/2 (box) and anti-targetABHD6. Of the other 20+ SH anti-targets, only modest (˜40%) inhibitionof MAGL was observed.

Example A9: Analysis of Cytotoxicity

Assay Overview: The purpose of this assay was to determine cytotoxicityof powder samples of test compounds. In this assay, HeLa cells in eitherserum-free medium or medium containing fetal calf serum (FCS) wereincubated with test compounds, followed by determination of cellviability. The assay utilized the WST-1 substrate, which was convertedinto colorimetric formazan dye by the metabolic activity of viablecells. The amount of formed formazan directly correlated to the numberof metabolically active cells in the culture. As designed, compoundsthat reduced cell viability resulted in decreased absorbance of the dye.

Protocol Summary: This assay was started by seeding HeLa cells in DMEMmedium supplemented with 10% FCS and 1× Pen-Strep-Glutamine in a 96-wellplate. Cells were incubated at 37° C. in a humidified incubator until80% confluent (24 hours). Medium was removed, and 100 μL of fresh medium(serum-free or supplemented with 10% FCS), pre-mixed with DMSO or testcompound, was added to each well. Cells were incubated for 48 hours at37° C. in a humidified incubator and cell viability was determined bythe WST-1 assay (Roche) according to manufacturer instructions. CC₅₀values were determined from dose-response curves from three replicatesat each inhibitor concentration (10, 100, 1000, 10000, 100000 nM),compared to six replicates of cells treated with DMSO only. AssayCutoff: Compounds with CC₅₀ values less than 10 μM were consideredactive (cytotoxic).

Compound 1 and analog Compound 2 were evaluated for cytotoxicity in HeLacells cultured in both serum-free and serum-supplemented medium.Compound 1 did not show evidence of cytotoxicity under either conditionup to 100 μM test concentration (see FIG. 5). The CC₅₀ for the˜equipotent but less selective Compound 2 appeared somewhat lower, witha lower limit of ˜10 μM. It should be noted that the solubility ofCompound 1 was determined to be 18 μM in medium alone and 46 μM inmedium supplemented with 10% serum. As such, the cytotoxicity resultsshould be interpreted with some caution. However, setting a lower CC₅₀limit equal 10 μM (highest test concentration below the determinedsolubility limit for Compound 1) still afforded a large dosing window(˜70-fold, based on in situ IC₅₀ values of 87/143 nM for LYPLA1/2) forinhibition of LYPLA1/2 without concern for overt toxicity in biologicalstudies.

Example A10: Gel Filtration to Assess Mode of Action

Assay Overview: The purpose of this assay was to determine whetherpowder samples of test compounds inhibit LYPLA1 and LYPLA2 in areversible or irreversible manner. In this assay, a complex proteome wasincubated with test compound and a fraction of the assay mixture waspassaged over a Sephadex G-25M column before reaction with FP-Rh andSDS-PAGE analysis as described above. As designed, test compounds thatacted as irreversible inhibitors prevented enzyme-probe interactionsboth before and after gel filtration, leading to low fluorescenceintensity in the band in the gel. In contrast, compounds that acted asreversible inhibitors showed recovery of probe labeling (and higherfluorescence intensity in the band in the gel) following gel filtrationto remove small molecules from the sample.

Protocol Summary: HeLa cell lysate (1 mL of 1 mg/ml in DPBS) was treatedwith test compound (200 nM or 1000 nM, from 50× stock in DMSO) or DMSOonly (no compound control). Test compounds were incubated for 30 minutesat 37° C. An aliquot (50 μL) was removed from each reaction, and theremaining sample was passaged over a Sephadex G-25M desalting column. Analiquot (50 μL, 1 mg/mL protein, eluted in DPBS) was removed, and pre-and post-filtration aliquots were reacted with FP-Rh (50× stock in DMSO;2 μM final concentration). The reaction was incubated for 30 minutes at25° C., quenched with 16 μL 4× SDS-PAGE loading buffer (reducing),separated by SDS-PAGE and visualized by in-gel fluorescent scanning. Thepercentage activity remaining was determined by measuring the integratedoptical density of the LYPLA1/2 bands (quantified together) relative toa DMSO-only (no compound) control. The percent recovery was calculatedby comparing percent inhibition before and after gel filtration for eachtest compound concentration.

An irreversible binding mode for Compound 1 was determined from gelfiltration studies, which demonstrated no recovery of ABPP probe FP-Rhlabeling for LYPLA1/2 following gel filtration to remove small molecules(e.g., inhibitor compounds), as shown in FIG. 6A.

Example A11: Serine Hydrolase (SH) Inhibition

Some N-hydroxy bicyclic hydantoin carbamates showed activity against oneor more brain SHs (see FIG. 7A-1).

Compound 5 was selected for analysis by mass spectrometry (MS)-basedABPP methods to assess the range of SHs targeted by N-Hydroxy bicyclichydantoin carbamates. Compound 5-sensitive SHs were identified using thequantitative MS method ABPP-SILAC (Stable isotope labeling by aminoacids in cell culture. Compound 5 was found to inhibit several human SHs(defined as proteins showing a three-fold or greater decrease in signalin the Compound 5-treated proteome), including enzymes previously shownto be sensitive to NHS-carbamates (e.g., ABHD6, MGLL) and others forwhich selective inhibitors are lacking (e.g., ABHD4, ABHD12, PLA2G15,PNPLA4) (see FIG. 7A-2).

A subset of N-hydroxy bicyclic hydantoin carbamates were selected forconcentration-dependent profiling, which identified compounds with goodpotency and selectivity for ABHD6(4-isopropyl-2,5-dioxoimidazolidin-1-yl4-(4-methoxyphenyl)piperazine-1-carboxylate, MJN193) and LYPLA1/2(Compound 1), as well as a promising lead inhibitor for ABHD3 and ABHD4(Compound 38) (see FIG. 7B).

Example A12: Optimization of N-Hydroxy Bicyclic Hydantoin CarbamateInhibitors for ABHD3 and ABHD4

Human ABHD3 and ABHD4 were recombinantly expressed by transienttransfection in HEK293T cells, and the cell lysates were combined toprovide a convenient assay for monitor inhibition of both enzymes bygel-based ABPP (see FIG. 8A). Gel-based ABPP confirmed that Compound 38acted as a potent dual inhibitor of ABHD3 and ABHD4 (IC₅₀ values of 0.13and 0.03 μM, respectively) and identified additional compounds thatpreferentially inhibited ABHD4 over ABHD3 (e.g., Compound 37, Compound36, and Compound 13) (see FIGS. 8B and 8C).

The activity and selectivity of dual ABHD3/4 and ABHD4-preferringinhibitors in PC3 cells by ABPP-SILAC was assessed. Isotopically heavyand light PC3 cells were treated in situ with inhibitor (Compound 37 (1μM) or Compound 38 (0.5 μM)) or DMSO for 4 h and then lysed andprocessed for ABPP-SILAC analysis. Compound 38 was found to inhibit bothABHD3 and ABHD4 in PC3 cells with good selectivity (see FIG. 9). Amongthe 44 serine hydrolases detected in this study, only four additionaltargets were observed for Compound 38—LIPE, PLA2G7, ABHD6, and CES2.Compound 37 completely inhibited ABHD4 and shared a similar off-targetprofile with Compound 38. Partial inhibition (˜80%) of ABHD3 was alsoobserved, which indicates that the selectivity window for ABHD4 overABHD3 may be compressed in living cells compared to cell lysates.

Example A13: Identification of PPT1 as a Target of N-Hydroxy BicyclicHydantoin Carbamates

Compound 39 inhibited ABHD4 in transfected HEK293T cell proteome in aconcentration-dependent manner as detected by competitive ABPP withFP-Rh, and the Compound 39-ABHD4 adduct was, in a complementary manner,directly visualized by CuAAC conjugation to a rhodamine-azide (Rh-N3)tag (see FIG. 10). Compound 39 also identified an Compound 37-sensitiveprotein in mouse brain that matches the expected molecular mass (˜40kDa) of ABHD4 and was absent in brain tissue from ABHD4−/− mice (seeFIG. 11). These data indicate that Compound 39 acts as a tailoredactivity-based probe for the convenient gel-based detection of ABHD4 incomplex biological systems.

Example A14: Competitive ABPP-SILAC Experiments to Identify Targets ofCompound 37 Using Compound 39 as a Probe

Isotopically heavy- and light-labeled PC3 cells were treated withCompound 37 (1 μM) or DMSO for 4 h, respectively, lysed, and thentreated Compound 39 (5 μM, 1 h). The heavy and light proteomic sampleswere then combined and analyzed by LC-MS/MS, which revealed a smallsubset of SHs, including ABHD4, that were inhibited by Compound 37 (seeFIG. 12). Among the handful of additional SH targets of Compound 37 wereABHD6 and PLA2G7, which were expected based on our ABPP-SILAC studieswith FP-biotin (see FIG. 9), and protein-palmitoyl thioesterase 1(PPT1), which was inhibited by more than 90%. PPT1 shows poor reactivitywith broad-spectrum FP probes. Accordingly, PPT1 was not detected in ourprevious ABPP-SILAC studies with FP-biotin (see FIG. 9). The selectiveenrichment of PPT1 by Compound 39 was confirmed by performing aprobe-versus-probe ABPP-SILAC study, where heavy and light PC3 proteomeswere treated with FP-biotin (2.5 μM, 1 h) and Compound 39 (5 μM, 1 h),respectively. PPT1, along with two additional SHs LIPA and DDHD2, werepreferentially enriched by Compound 39 over FP-biotin, while most of theother SHs were more strongly enriched by FP-biotin (see FIG. 13). Ahandful of SHs corresponding mostly to enzymes that were identified inour ABPP-SILAC studies as targets of Compound 37 (see FIGS. 9 and 12),were equivalently enriched by Compound 39 and FP-biotin (see FIG. 13).

The selective enrichment of PPT1 by Compound 39 and the near-completeblockade of this enrichment by Compound 37 suggested that this enzymewas covalently modified and inhibited by N-hydroxy bicyclic hydantoincarbamates. This possibility was explored by treating HEK293T cellstransfected with human PPT1 (hPPT1) with Compound 37 (5 μM, 4 hpre-treatment) or DMSO followed by cell lysis and treatment withCompound 39 (5 μM, 1 h). A strong, but diffuse Compound 37-sensitive,Compound 39-labeled protein band was detected by gel-based ABPP inhPPT1-transfected, but not mock-transfected cell lysates (see FIG. 14),and this diffuse signal was compressed to a tight, faster-migrating bandmatching the predicted molecular weight of PPT1 following treatment withthe glycosidase PNGaseF (see FIG. 14). Western blotting with ananti-PPT1 antibody confirmed a similar migration pattern for hPPT1protein in transfected cell lysates (see FIG. 14, bottom panel). Thesedata indicated that both Compound 37 and Compound 39 reacted with PPT1and are consistent with previous studies demonstrating that this enzymeis glycosylated. In contrast, FP-Rh failed to detect hPPT1 intransfected cell lysates (see FIG. 14).

Example A15: Reaction of N-Hydroxy Bicyclic Hydantoin Carbamates withthe Catalytic Serine of PPT1-S115

HEK293T cells were transfected with wild type and an S115A mutant ofPPT1, lysed, and treated with Compound 39 followed by CuAAC conjugationto an azide-Rh tag, PNGaseF treatment, and analysis by SDS-PAGE. Thisstudy revealed strong labeling of wild-type PPT1, but not the S115Amutant (see FIG. 15). S115 was identified as the site of Compound 39labeling on PPT1 using a quantitative MS method termed isoTOP-ABPP. Inbrief, PPT1-transfected cell lysates were treated with DMSO or theCompound 39 probe (5 μM) for 1 h and then incubated with isotopicallyheavy or light azide-biotin tags, respectively, under CuAAC reactionconditions. Each azide biotin tag also contained an intervening TEVprotease cleavage site. Heavy and light probe-labeled proteins were thencombined, enriched by streptavidin chromatography, digested sequentiallyon-bead with trypsin (to remove non-probe-modified peptides) and TEV torelease Compound 39-modified peptide(s), which were analyzed by LC-MS/MSon an LTQ-Orbitrap instrument. A single Compound 39-modified peptide,corresponding to amino acids 105-122, was detected in the isotopicallylight, but not heavy samples, and tandem MS analysis assigned theCompound 39 modification site to S115 (see FIG. 16). Taken together,these data indicated that N-hydroxy bicyclic hydantoin carbamates reactwith PPT1 through carbamylation of the enzyme's conserved serinenucleophile.

Example A16: Selective Inhibitors for ABHD4 and PPT1 Enzymes

A structurally diverse panel of N-hydroxy bicyclic hydantoin carbamateswere assayed at 2 or 0.2 μM for in situ inhibition of hPPT1 intransfected HEK293T cells. While many compounds exhibited inhibitoryactivity toward PPT1 at 2 μM, only Compound 26 showed completeinhibition of hPPT1 at 200 nM. Compound 26 displayed an in situ IC₅₀value for inhibiting recombinant PPT1 of 0.1 μM as measured by gel-basedcompetitive ABPP (Table 7).

Table 7 shows IC₅₀ values (μM; with 95% confidence intervals inparentheses) for inhibition of human PPT1 by Compounds 1, measured inPPT1-transfected cells (in situ, competitive ABPP) or cell lysates (invitro, pH 5.0, MU-65-Palm-βD-Glc substrate).

Cpd. in situ (ABPP) in vitro (substrate) 26 0.1 (0.04-0.2) 6.5 (3.6-12)34 3.9 (2.0-7.4) >50 1 0.5 (0.3-1.0)  15 (9.5-25)

Compound 26 was then tested to see if it inhibited PPT1 activity using asubstrate assay. Endogenous substrates for PPT1 remain poorlycharacterized, but this enzyme was assayed with a fluorogenic substrate4-methylumbelliferyl-6-thio-palmitate-βD-glucopyranoside(MU-6S-Palm-βD-Glc). In this assay, PPT1 hydrolyzes the substrate'spalmitoyl thioester bond, and exogenous sweet almond β-glucosidasedeglycosylates and frees the fluorophore, 4-methylumbelliferone (4-MU),which is measured to assess PPT1 activity. PPT1-transfected cellsdisplayed a strong increase in MU-6S-Palm-βD-Glc hydrolysis compared tomock-transfected cells (see FIG. 17A), and treatment of thesetransfected lysates at pH 5.0 with Compound 26 fully blocked PPT1activity with an IC₅₀ value of 6.5 μM (see FIG. 17B and Table 7).Additionally, in situ treatment of PC3 cells with Compound 26 (500 nM, 4h) showed a 90% reduction in endogenous PPT1 activity based on theMU-6S-Palm-βD-Glc substrate assay (see FIG. 17C).

Compound 34, which showed limited activity against PPT1 in situ (seeFIG. 17D and Table 7) or in vitro (see FIG. 17A and Table 7), was a goodinhibitor of ABHD4 in vitro (IC₅₀ value of 0.5 μM) as measured with anN-acyl phosphatidylethanolamine (NAPE) substrate hydrolysis assayperformed with lysates from ABHD4-transfected HEK293T cells (see FIG.18). In contrast, neither Compound 26 nor Compound 1 altered the NAPEhydrolysis activity of ABHD4. Compound 1 did, however, inhibit PPT1activity both in vitro (see FIG. 17B and Table 7) and in situ (Table 7),albeit with less potency than Compound 26.

To evaluate the broader selectivity of Compound 26 and Compound 34across the SH class, heavy and light isotopically-labeled PC3 cells weretreated with inhibitor [Compound 34 (1 μM); or Compound 26 (1 or 0.1μM); 4 h) and DMSO, respectively, followed by ABPP-SILAC analysis witheither the FP-biotin probe or the Compound 39 probe. Compound 26, testedat 1 or 0.1 μM, near-completely inhibited PPT1 (˜90%+; as measured withthe Compound 39 probe) and showed good selectivity, only cross-reactingwith two of the >40 quantified SH activities—ABHD6 and CPVL (see FIGS.19 and 20). Conversely, Compound 34 inhibited >90% of ABHD4 activity, asmeasured with either the FP-biotin probe or the Compound 39 probe, andcross-reacted with four additional targets—ABHD6, PAFAH2, PLA2G7, andPLA2G15.

These results, taken together, demonstrate that Compound 26 and Compound34 were cell-active inhibitors that displayed good potency andselectivity for PPT1 and ABHD4, respectively.

Example A17: In Vivo Activity

Compound 26, Compound 34, and Compound 1 were administered across a doserange of 1-40 mg/kg to C57BL6 mice by intraperitoneal injection, and,after 4 hours, animals were sacrificed and tissues harvested foranalysis by gel-based ABPP using the FP-Rh probe and the Compound 39probe. Compound 26 showed potent, dose-dependent inhibition of anCompound 39-reactive protein matching the molecular mass ofdeglycosylated PPT1 in PNGaseF-treated central and peripheral tissues,producing near-complete blockade of this enzyme activity at 1-5 mg/kg(see FIGS. 21 and 22). Compound 26 showed excellent selectivity for PPT1and did not inhibit other SHs detected in brain and testis tissuestreated with the FP-Rh probe (see FIG. 22) or the Compound 39 probe (seeFIGS. 21 and 22). Compound 1 also showed good in vivo activity,producing near-complete blockade of LYPLA1 and LYPLA2 in peripheral(e.g., liver, kidney), but not central (brain) tissues at a dose of 5mg/kg as measured with the FP-Rh probe (FIG. 23). Compound 1 also showedlimited cross-reactivity with other SHs, consistent with competitiveABPP-SILAC studies in human cells (FIG. 24) but did inhibit PPT1 atdoses of 10 mg/kg or greater as measured with the Compound 39 probe.Contrasting with these results, Compound 34 did not block the activityof ABHD4 in any tissue examined, possibly indicating that improvementsin potency and/or drug-like properties are required to convert thiscompound into an in vivo-active probe.

We claim:
 1. A compound, or a stereoisomer or a pharmaceuticallyacceptable salt thereof, of formula (I):

wherein W is

each m is independently 0, 1, or 2; R¹ is H, halo, —OH, cyano, amino,(C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₆-C₁₀)aryl, or (C₆-C₁₀)aryl(C₁-C₈)alkyl;R² is H, —COOR⁵, or —CONR⁵R⁶; R³ is H, (C₆-C₁₀)aryl, 5-9 memberedheteroaryl comprising 1 or 2 heteroatoms independently selected from O,N, and S, (C₆-C₁₀)aryl(C₁-C₈)alkyl, or 5-9-memberedheteroaryl(C₁-C₈)alkyl comprising 1 or 2 heteroatoms independentlyselected from O, N, and S; wherein (C₆-C₁₀)aryl, 5-9-memberedheteroaryl, (C₆-C₁₀)aryl(C₁-C₈)alkyl, and 5-9-memberedheteroaryl(C₁-C₈)alkyl are optionally substituted by one or moresubstituents independently selected from halo, (C₁-C₈)alkyl,(C₁-C₈)alkoxy, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₂-C₈)alkenyloxy,(C₂-C₈)alkynyloxy, —C(O)OR^(a), and —C(O)N(R^(a))₂; each R^(a) isindependently hydrogen, (C₁-C₈)alkyl, or (C₆-C₁₀)aryl; R⁴ is(C₆-C₁₀)aryl, 5-9 membered heteroaryl comprising 1 or 2 heteroatomsindependently selected from O, N, and S, (C₆-C₁₀)aryl(C₁-C₈)alkyl,5-9-membered heteroaryl(C₁-C₈)alkyl comprising 1 or 2 heteroatomsindependently selected from O, N, and S, or di(C₆-C₁₀)aryl(C₁-C₈)alkyl;wherein (C₆-C₁₀)aryl, 5-9-membered heteroaryl, (C₆-C₁₀)aryl(C₁-C₈)alkyl,and 5-9-membered heteroaryl(C₁-C₈)alkyl are optionally substituted byone or more substituents independently selected from (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, halo, (C₁-C₈)fluoroalkyl, —OR^(a), and(C₆-C₁₀)aryl optionally substituted with one or more halo groups; R⁵ isH or (C₁-C₈)alkyl; R⁶ is (C₆-C₁₀)aryl, 5-9 membered heteroarylcomprising 1 or 2 heteroatoms independently selected from O, N, and S,(C₆-C₁₀)aryl(C₁-C₈)alkyl, or 5-9-membered heteroaryl(C₁-C₈)alkylcomprising 1 or 2 heteroatoms independently selected from O, N, and S;and R′ is H, (C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₁-C₈)alkylcarbonyl,(C₁-C₈)alkoxycarbonyl, (C₃-C₉)cycloalkyl, (C₃-C₉)cycloalkyl(C₁-C₈)alkyl,(C₃-C₉)cycloalkylcarbonyl, (C₃-C₉)cycloalkoxycarbonyl, (C₆-C₁₀)aryl,(C₆-C₁₀)arylcarbonyl, (C₆-C₁₀)aryloxycarbonyl, (C₆-C₁₀)aryl(C₁-C₈)alkyl,(C₆-C₁₀)aryl(C₁-C₈)alkylcarbonyl, (C₆-C₁₀)aryl(C₁-C₈)alkoxycarbonyl, 5-9membered heterocyclyl comprising 1 or 2 heteroatoms independentlyselected from O, N, and S, 5-9 membered heterocyclyl(C₁-C₈)alkylcomprising 1 or 2 heteroatoms independently selected from O, N, and S,5-9-membered heterocyclylcarbonyl comprising 1 or 2 heteroatomsindependently selected from O, N, and S, 5-9 membered heteroarylcomprising 1 or 2 heteroatoms independently selected from O, N, and S,5-9 membered heteroaryl(C₁-C₈)alkyl comprising 1 or 2 heteroatomsindependently selected from O, N, and S, or 5-9-memberedheteroarylcarbonyl comprising 1 or 2 heteroatoms independently selectedfrom O, N, and S; wherein (C₆-C₁₀)arylcarbonyl,(C₆-C₁₀)aryl(C₁-C₈)alkyl, and 5-9-membered heterocyclyl(C₁-C₈)alkyl areoptionally substituted by one or more substituents independentlyselected from (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, halo,(C₁-C₈)fluoroalkyl, nitro, (C₆-C₁₀)aryl(C₁-C₈)alkyl, —OR^(a),—N(R^(a))₂, —C(O)OR^(a), and —C(O)N(R^(a))₂.
 2. The compound of claim 1,or a stereoisomer or a pharmaceutically acceptable salt thereof, whereineach m is
 1. 3. The compound of claim 2, or a stereoisomer or apharmaceutically acceptable salt thereof, wherein R¹ is H.
 4. Thecompound of claim 3, or a stereoisomer or a pharmaceutically acceptablesalt thereof, wherein W is


5. The compound of claim 4, or a stereoisomer or a pharmaceuticallyacceptable salt thereof, wherein: R² is H; and R³ is (C₆-C₁₀)aryl or(C₆-C₁₀)aryl(C₁-C₈)alkyl; and R³ is optionally substituted by one ormore substituents independently selected from halo, (C₁-C₈)alkyl,(C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl, (C₂-C₆)alkenyloxy,(C₂-C₆)alkynyloxy, —C(O)OR^(a), and —C(O)N(R^(a))₂.
 6. The compound ofclaim 5, or a stereoisomer or a pharmaceutically acceptable saltthereof, wherein R³ is unsubstituted phenyl, unsubstituted benzyl, orunsubstituted phenethyl.
 7. The compound of claim 5, or a stereoisomeror a pharmaceutically acceptable salt thereof, wherein R³ is phenyl,benzyl, or phenethyl, and R³ is optionally substituted with one or moresubstituents independently selected from the group consisting of halo,(C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,(C₂-C₆)alkenyloxy, and (C₂-C₆)alkynyloxy.
 8. The compound of claim 7, ora stereoisomer or a pharmaceutically acceptable salt thereof, wherein R³is phenyl or phenethyl, and R³ is substituted with one substituentselected from the group consisting of fluoro, chloro, bromo, methoxy,ethynyl, and propargyloxy.
 9. The compound of claim 3, or a stereoisomeror a pharmaceutically acceptable salt thereof, wherein W is


10. The compound of claim 9, or a stereoisomer or a pharmaceuticallyacceptable salt thereof, wherein R⁴ is (C₆-C₁₀)aryl or(C₆-C₁₀)aryl(C₁-C₈)alkyl; and R⁴ is optionally substituted by one ormore substituents independently selected from halo, (C₁-C₈)alkyl,(C₁-C₈)alkoxy, (C₂-C₆)alkenyl, and (C₂-C₆)alkynyl.
 11. The compound ofclaim 10, or a stereoisomer or a pharmaceutically acceptable saltthereof, wherein R⁴ is unsubstituted phenyl, unsubstituted benzyl, orunsubstituted phenethyl.
 12. The compound of claim 10, or a stereoisomeror a pharmaceutically acceptable salt thereof, wherein R⁴ is phenyl,benzyl, or phenethyl, and R⁴ is optionally substituted with one or moresubstituents independently selected from the group consisting of halo,(C₁-C₈)alkyl, (C₁-C₈)alkoxy, (C₂-C₆)alkenyl, and (C₂-C₆)alkynyl.
 13. Thecompound of claim 12, or a stereoisomer or a pharmaceutically acceptablesalt thereof, wherein R⁴ is phenyl or phenethyl, and R⁴ is substitutedwith one substituent selected from the group consisting of fluoro,chloro, bromo, methoxy, and ethynyl.
 14. The compound of claim 3, or astereoisomer or a pharmaceutically acceptable salt thereof, wherein R′is H, unsubstituted (C₁-C₈)alkyl, unsubstituted (C₁-C₈)alkylcarbonyl,unsubstituted (C₁-C₈)alkoxycarbonyl, unsubstituted (C₆-C₁₀)arylcarbonyl,unsubstituted (C₆-C₁₀)aryl(C₁-C₈)alkyl, unsubstituted 5-9-memberedheterocyclyl(C₁-C₈)alkyl comprising 1 or 2 heteroatoms independentlyselected from O, N, and S, or unsubstituted 5-9-memberedheterocyclylcarbonyl comprising 1 or 2 heteroatoms independentlyselected from O, N, and S.
 15. The compound of claim 3, or astereoisomer or a pharmaceutically acceptable salt thereof, wherein: R′is (C₁-C₈)alkylcarbonyl, (C₆-C₁₀)aryl(C₁-C₈)alkyl, or 5-9 memberedheterocyclyl(C₁-C₈)alkyl comprising 1 or 2 heteroatoms independentlyselected from O, N, and S; wherein (C₆-C₁₀)arylcarbonyl,(C₆-C₁₀)aryl(C₁-C₈)alkyl, and 5-9-membered heterocyclyl(C₁-C₈)alkyl areoptionally substituted by one or more substituents independentlyselected from (C₁-C₈)alkyl halo, nitro, (C₆-C₁₀)aryl(C₁-C₈)alkyl —OR^(a)and —N(R^(a))₂.
 16. The compound of claim 14, or a stereoisomer or apharmaceutically acceptable salt thereof, wherein R′ is unsubstituted5-9-membered heterocyclylcarbonyl comprising 1 or 2 heteroatomsindependently selected from O, N, and S.
 17. The compound of claim 16,or a stereoisomer or a pharmaceutically acceptable salt thereof,wherein: R′ is


18. A compound, or a stereoisomer or a pharmaceutically acceptable saltthereof, selected from:


19. A pharmaceutical composition comprising a compound of claim 1 and atleast one pharmaceutically acceptable excipient.
 20. A method oftreatment of a medical condition in a patient comprising administeringan effective dose of a compound of claim 1 to the patient, wherein themedical condition is selected from pain, epilepsy, and traumatic braininjury.